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Coombe & McCarthy Grape berry growth & ripening 131

Dynamics of grape berry growth and physiology of ripening

B.G. COOMBE1 and M.G. McCARTHY2,3
1
Department of Horticulture,Viticulture and Oenology,Waite Campus,The University of Adelaide,
Glen Osmond, SA 5064
2
Nuriootpa Viticulture Centre, PO Box 245, Nuriootpa SA 5355,Australia
3
Corresponding author: Dr M. McCarthy, facsimile +61 8 8568 6449, e-mail mccarthy.michael@saugov.sa.gov.au

Abstract
Data from two experiments on development of grape berries is re-examined with emphasis on
partitioning of berry weight into non-solutes per berry (largely water) and solutes per berry (largely
sugar), using weight times juice °Brix. This approach is based on the thought that, since xylem flow is
blocked after veraison, time curves of solutes per berry indicate the activity of phloem transport into the
berry during ripening growth. Experiment 1: Measurements of Muscat Gordo Blanco berries from
inflorescences with a spread of flowering times showed typical double-sigmoid volume/time curves but
with divergent rates and amounts of volume increase. Despite this divergence, °Brix curves after veraison
were almost coincident because, in each case, the rate of increase in solutes per berry was proportional to
that of berry volume. These results indicate that sugar and water increments after veraison are linked and
depend on the same source, namely, phloem sap. Experiment 2: An irrigation experiment on cv. Shiraz also
showed divergent berry weight curves between treatments and years but with the difference that all
berries shrank after a maximum berry weight was attained at 91 days after flowering (at about 20 °Brix).
At this point, the curves of solutes per berry slowed then plateaued, indicating that inflow of phloem sap
had become impeded. Prior to shrinkage these berries accumulated primary metabolites (mainly phloem
sugar) but, during shrinkage, when berries were apparently isolated from vascular transport, non-
anthocyanin glycosides accumulated. These results have implications for the study of berry flavour build-
up and berry composition, and also for the understanding of sink competition within the vine, fresh and
dried yield, and juice °Brix levels.

Keywords: grape berry, berry weight, berry solutes, berry water, xylem and phloem transport

Introduction In this review, the interplay of xylem and phloem

The growth of a grape berry consists of two successive sig-
moid cycles, each with distinctive characteristics (Coombe
1992). The first cycle—berry formation—begins with a
spate of cell division in pericarp tissue the amount and
direction of which largely determines a berry’s final size
and shape. The rate of these divisions is positively corre-
lated with the growth rates of developing seeds in each
berry. Pericarp cell division changes gradually to cell
enlargement, which later slows as the first sigmoid cycle
ends. At this stage the berry is hard, green and slow-
growing; the principal organic compound accumulating is
malic acid.
The second cycle begins with the onset of sugar accu-
mulation, berry softening, berry colouring, and renewed
size increase; taken collectively these events constitute
veraison, denoting the beginning of ripening processes.
Ripening is characterised by the accumulation in flesh
and skin of hexose sugars and in skin of potassium and
phenolics (including anthocyanins in black grapes)
(Coombe 1987). Flavour compounds accumulate late in
ripening, a stage that has been termed ‘engustment’ by
Coombe and McCarthy (1997).
transport into the grape berry and of metabolism within
the berry are examined with respect to the determination
of berry composition.
Increments of water and sugar
A seminal experiment has been reported (Coombe 1980)
in which the development of grape berries was exam-
ined by separating the two chief components of berry
weight: non-solutes per berry (largely water) and solutes
per berry (largely sugar), their ratio being concentration
e.g. °Brix. Inflorescences on Muscat Gordo Blanco vines
were induced to develop at different times by a combina-
tion of pruning and topping treatments, making possible
the selection of groups with a spread of six flowering
times. On each of these six groups of bunches, berries
were marked 12 days after 70% capfall and their dia-
meters measured weekly until maturity. From the lag-
phase onwards, single berries were sampled from each
bunch for measurement of juice °Brix. Calculated berry
volumes (Figure 1a) showed divergent double-sigmoid
time curves with nearly a two-fold range of lag-phase
volumes; in most cases these differences in rates and
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132 Grape berry growth & ripening Australian Journal of Grape and Wine Research 6, 131–135, 2000

Impedance of xylem transport at veraison

The above observations raise questions about the contri-
3 bution of xylem transport to water movement into
berries during both major phases of development. Xylem
A sap is a dilute aqueous solution of inorganic ions and
some root-derived organic metabolites. Greenspan et al.
Berry volume (mL)

2 (1994) have shown that xylem sap is the main source of

water for berries during their first growth cycle during
which dry matter accumulation is relatively small.
25 However, when berries resume swelling at the beginning
1 of the second growth cycle, the flow of xylem sap into
20 the berry becomes impeded. This was shown in two con-
current papers using different methods: Düring et al.
15 (1987) found, using two staining materials, that breaks

°Brix
0 developed throughout the peripheral bundle system after
10 berries had started to expand during the ripening cycle;
B Findlay et al. (1987) perfused 14C-sucrose or eosin dye
5 through the cut pedicel of detached berries and found
0.6 that, after veraison, xylem flow into flesh was blocked
Solutes per berry (g)

0 due to stretched and broken segments of tracheids,

principally those at the top of the pedicel (the brush).
0.4 This phenomenon was confirmed in two other varieties
by Creasy et al. (1983).
This disorder appears to be due to stretching of
0.2 C
tracheids and breakage of tracheid wall membranes,
especially in the brush zone where the vascular bundles
0 enter the berry. Subsequent berry growth would then
0 50 100 150 depend mainly on the flow of phloem sap, thus support-
Days after flowering ing the previous explanations of the linkage between
water and sugar increments during ripening growth. That
Figure 1. Changes in (a) berry volume, (b) juice °Brix, and (c) the explanation is feasible is supported by calculations
weight of solutes per berry against days after flowering of cv.
Muscat Gordo Blanco berries on bunches that flowered at weeks 1, using a notional percentage of sucrose in phloem sap and
2, 4, 5, 6 and 7 beginning with week 1 on 1 November (after the weights of sugar and water that accumulate during
Coombe 1980). ripening. The suggestions are supported by the research
of Greenspan et al. (1994).

Impedance of phloem transport?

amount of volume increase were maintained until har-
vest. Despite these differences, curves of juice °Brix were
parallel (Figure 1b); when this figure is redrafted with
the x-axis as ‘days from veraison’ the Brix curves are
practically coincident (see Figure 1 of Coombe and Iland
1987). This coincidence was because, in all cases, the
increases in solutes per berry (Figure 1c) were propor-
tional to the divergent rates of increase in berry volume
during most of the ripening growth cycle.
Recognising that sugar is the predominant component
of berry solutes, and water of volume, these results indi-
cate that sugar and water increments were linked to the
one source. The most probable source is phloem sap. The
fact that the curves of solutes per berry increased
throughout indicates that phloem transport was un-
impeded. Subsequent work (Coombe and Phillips 1982)
has emphasised the nexus between water and sugar
increments, and hence the regularity of concentration
increases. Evidence for this conclusion was also found
with cv. Dolcetto in Italy by Coombe et al. (1987), who
concluded that the primary control of accumulation of
both solutes and non-solutes was unloading of phloem
sap into berry flesh.
Although not dealing with mechanisms, the foregoing is
an attempt to explain the logistics concerning water and
sugar accumulation in grape berries. A reason for modifi-
cation of this explanation, at least for some varieties
such as Shiraz, has emerged from the results of a field
experiment at Waikerie by McCarthy (1997a, 1997b)
comparing eight irrigation treatments with nine repli-
cates over four years. Berry weight/time curves showed
an array of different dimensions attributable to treat-
ments and years, with all following a typical double-
sigmoid shape for most of their development. However,
in all cases, the berry weight curves showed a decrease
beginning midway through the second cycle at 91 ± 3
days after flowering (McCarthy 1999). The dimensions of
berry shrinkage are illustrated in Figure 2a.
Partition of berry weight during the second cycle into
non-solutes per berry (mainly water) and solutes per
berry (mainly sugar) showed that curves of non-solutes
per berry matched those of berry weight in general shape
and with the same timing of maximum berry weight
(Figure 2c). Curves of solutes per berry (Figure 2d) dif-
fered from those of non-solutes per berry in showing an
initial rapid rise, typical of the stage of rapid influx of
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Coombe & McCarthy Grape berry growth & ripening 133

2.0 B 25 to the continuation of loss of water by transpiration, thus

causing berry shrinkage (see references in McCarthy and
Coombe 1999).
What could cause blockage of phloem transport?
20
Using 2-year-old grapevine canes, Esau (1948) showed
that blockage of stem phloem by the deposition of callose
1.5

°Brix
plugs on sieve areas was a normal occurrence during
15 autumn, and that plugging also occurred at other times
during summer. A case for investigation of this phenom-
Grams

enon in relation to shrinkage of Shiraz grape berries is

A 10 indicated. Such investigation should take account of the
1.0 intriguing aspects related to the incidence of shrinkage
shown by McCarthy (1999), namely, that timing of
C shrinkage onset was more closely related to the number
5 of days after flowering than to environmental factors
0.3 such as soil water stress, rainfall or heat summation. The
D
results imply operation of a developmentally-controlled
0.2
timing mechanism associated with functioning of phloem
0.1 elements. A systemic phenomenon is indicated, but the
anatomical location of the blockage is unclear.
0.0
60 70 80 90 100 110 120 130 Implications of phloem blockage
Days after flowering The proposed impedance of phloem transport into the
berry is a novel hypothesis which has important implica-
tions for the understanding of development in berries
Figure 2. Changes in four variables during ripening of berries that shrink (such as Shiraz) and hence of all berries.
sampled from an irrigation experiment on cv. Shiraz at Waikerie, Some are listed here.
South Australia: (a) weight per berry ■, (b) juice °Brix ●, (c) non-
solutes per berry ▲, and (d) solutes per berry ▼. Data from
McCarthy (1997a, 1997b, 1999) and McCarthy and Coombe (1999);
each point is an average of nine replicates and seven treatments
from year 4 harvest.
300

Solutes
Milligrams

per berry
200

1.0
100
µmole

0.5

Figure 3. Hypotheses from Figure 2 (from McCarthy and Coombe

1999, reproduced with permission).

Red-free G-G
sugar, then a distinct slowing in the rate of increase co- per berry
incident with the beginning of berry shrinkage; after
0.0
another 2–3 weeks solute increase per berry had stopped 10 15 20 25
(McCarthy and Coombe 1999). This is interpreted in °Brix
Figure 3 as indicating that, at maximum berry weight
(weight-max.), flow of phloem sap became impeded, and Figure 4. Weight of solutes per berry and non-anthocyanin
finally blocked 2–3 weeks later; thereafter, it would glucosides per berry (red-free G-G, as glucose equivalents) against
appear that at this time berries became isolated from both juice °Brix during ripening of Shiraz berries in year 4 of the fully-
irrigated treatment of an irrigation experiment at Waikerie, South
long-distance vascular transport pathways, i.e. xylem and Australia (McCarthy 1997a, 1997b). The points in the lower graph
phloem. The fact that non-solutes per berry declined derive from seven groups of five values with ascending °Brix levels
while solutes per berry stayed constant can be attributed and show the means and SE bars for the x and y axes.
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134 Grape berry growth & ripening Australian Journal of Grape and Wine Research 6, 131–135, 2000

1. Removal of fruit’s sink for photosynthate and reserves xylem

would permit flow to other sinks in the vine to an Vascular
extent depending on the timing of blockage and its flow normal
anatomical location. into berry
2. During shrinkage of Shiraz berries, yields are decreas- phloem shrinkage
ing and the juice °Brix increasing, with the latter
flavour
dependent on the loss of water during shrinkage since compounds
there is little further increase in fruit solutes. sugar

3. Such berries would appear to undergo engustment anthocyanin

processes in isolation from the rest of the vine
(Coombe and McCarthy 1997). Indeed, as Figure 4
shows, Shiraz berries had completed most of the
Berry formation
accumulation of solutes per berry by 20°Brix but all of
the increase in non-anthocyanin glycosides (red-free
G-G) occurred after that stage. malate

The metabolic picture indicated in (3) may not be

Berry veraison
restricted to varieties such as Shiraz that show berry volume
shrinkage. For instance, normally-expanding Muscat
berries (like those in Figure 1a) also showed increases in Berry ripening
pericarp
red-free G-G late in ripening (Gholami et al. 1996). This cell
division
is also evident in the results for several winegrape
varieties currently being analysed as part of the National setting
Vineyard Fruit Composition Survey (I.L. Francis, person-
al communication).
Further support for the notion that grape berries can 0 20
Days
40 60 80 100 120
undergo engustment (i.e. develop flavour) while isolated °Brix 4 7
Flowering 10 14 18 22 26
from phloem translocation is provided by the research of
Gholami (1996). By grafting inflorescences of varieties
that develop aromatic berry flavour onto vines that have Figure 5. Notional diagrams showing, at top, estimates of rates of
neutral berry flavour, and vice versa, it was shown that vascular flow into berries, partitioned between xylem and phloem,
the ripening berries in both cases developed a mono- their sum matching the rates of water input as shown by the berry
terpene glucoside analysis typical of their genotype with- volume growth curves below of Muscat (solid lines) and Shiraz
(dashed lines). The berry volume is an idealised curve against days
out influence from the genotype of the scion (Gholami et after flowering and juice °Brix and shows the berry shrinkage and
al. 1995). phloem blockage of Shiraz starting at about 18-20°Brix; the arrow-
heads at top signify the timing of the proposed impedance of xylem
Integration flow (in all varieties) and of phloem flow in berries that shrink. Three
A notional picture of the relative contributions of xylem successive but overlapping phases of solute accumulation are
indicated—malate, sugar and flavour compounds—each sequence
and phloem translocation to water and solute accumula- occurring in both varieties. Based on data in Coombe (1987, 1992),
tion by grape berries is given in Figure 5, integrating Coombe and McCarthy (1997), McCarthy (1997b) and McCarthy
Muscat and Shiraz results. Three phases are apparent. and Coombe (1999).

1. From setting to veraison

Berry volume (largely determined by the amount of cell
division in the pericarp during the first three weeks)
increases sigmoidally to the lag phase at which time the
berries remain hard and green. The water for expansion
during this phase derives from both xylem and phloem
transport, but especially xylem. The principal solute that
accumulates is malate.
2. From veraison to 18–20°Brix
With the onset of ripening in the second cycle, berries
soften and expand. Xylem flow is impeded, hence the
water that moves in during this phase is largely from
phloem sap along with sugar which is the chief solute
accumulating in flesh. Sugar and potassium accumulate
in skin cells, along with anthocyanins in coloured vari-
eties.
3. From 18–20°Brix to harvest
Development of berries during this phase differs between
the two varieties. Shiraz berries at Waikerie decreased in
weight. We have suggested this decrease was due to a
blockage of phloem transport into the berries so that the
water and sugar supplied by phloem sap was cut off—see
the dashed lines in Figure 5 and the plateau in solutes per
berry in Figure 2d. Volume decline was due to transpira-
tion losses with a consequential increase in juice °Brix.
Note that, on this interpretation, these berries during this
phase had little vascular connection to the rest of the
vine. With Muscat (solid lines in Figure 5), berry weights
continued their sigmoid increase and solutes per berry
did not depart from linear increases throughout (Figure
1a and 1c). These results imply that phloem sap supply
continued throughout ripening. The amounts of phloem

Coombe & McCarthy
supply are indicated by the increases in berry volume
and the differences in the slopes of the volume and solute
curves are explained by transpirational losses, as with
Shiraz. Another resemblance to Shiraz is that Muscat
berries have been shown to accumulate glucosides late in
ripening (Gholami et al. 1996). That is, accumulation of
non-anthocyanin glucosides in berries of these two
varieties proceeded with or without the benefit of phloem
sap input, indicating that input from phloem sap is not a
requirement for accumulation of such glucosides. This
conclusion was arrived at in a different way by Gholami
et al. (1995), who found by bunch grafting experiments
that accumulation of glucosides of aroma compounds was
determined by the genotype of the grafted bunch, not
the scion tissues.
The ideas raised here provoke a large number of
questions which deserve further research, for example: is
phloem blockage a reality and, if so, where, when and
why? If callose plugs sieve areas in phloem elements,
what are the conditions for callose formation? What are
the mechanisms controlling synthesis of flavour com-
pounds and their glucosides within berries? Why does
the synthesis of malvidin-3-glucoside appear linked to
sugar accumulation yet both precede synthesis of non-
anthocyanin glucosides? These and many other questions
are amenable to investigation using present physiological,
biochemical and molecular methods. Answers to these
and related questions should improve understanding of
the control of berry composition and especially flavour.
Acknowledgements
This review is a speculative integration of research by
graduate students and staff at the University of Adelaide
dealing with grape berry development. The authors are
indebted to Dr Robyn van Heeswijck and Dr Peter May,
Department of Horticulture, Viticulture and Oenology,
University of Adelaide, for constructive comments on the
manuscript.
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Manuscript received: 16 August 1999
Amended version received: 3 April 2000