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Scientia Horticulturae
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / s c i h o r t i
a r t i c l e i n f o a b s t r a c t
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Blueberries are now the second most economically important soft fruit. However, they are highly per-
Received 5 January 2015
ishable and susceptible to rapid spoilage. One of the main factors limiting postharvest life of
Received in revised form 11 March 2015
Accepted 13 March
blueberries is softening. The changes of fruit firmness, cell wall degrading enzymes and cell wall
2015 composition of
Available online 3 April 2015 ‘Brilliant’ blueberry (Vaccinium ashei cv. Brilliant) were investigated in this study. The results showed
fruit firmness declined concomitantly with the increase of the content of water soluble pectin (WSP)
Keywords: during storage paralleled by a decreasing amount of sodium carbonate soluble pectin (SSP), cellulose
Blueberry and hemicellulose. Blueberries stored at low temperature (5 ◦ C) maintained higher fruit firmness
Firmness than those stored at 10 ◦ C, which was due to the lower WSP content and higher contents of SSP,
Cell wall composition cellulose and hemicellulose. Meanwhile, the lower activities of cell wall degrading enzymes such as
Cell wall degrading enzymes
polygalacturonase, cellulase, -galactosidase and -mannosidase in blueberries at 5 ◦ C were
associated with greater fruit firmness and lower WSP content as compared to those in fruit stored at
10 ◦ C.
© 2015 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.scienta.2015.03.018
0304-4238/© 2015 Elsevier B.V. All rights reserved.
H. Chen et al. / Scientia Horticulturae 188 (2015) 44–48 45
46 H. Chen et al. / Scientia Horticulturae 188 (2015) 44–48
2.5 three pectin-rich fractions as WSP, CSP, and SSP and hemicellulose
and cellulose. Among them, WSP was thought to represent wall
o
5 C polymers solubilized in vivo but remaining in the apoplast which
2.0 10 oC could be obtained by water extraction of the purified cell wall,
whereas CSP and SSP are generally considered to be enriched for
* * * ionically and covalently bound pectins, respectively. Typically, dur-
ing fruit softening, depolymerisation of cell wall polysaccharides
Firmness (N)
1.5 * was accompanied by increases in the levels of WSP, whilst the lev-
*
els of CSP, SSP, hemicellulose and cellulose decreased (Carrington
* et al., 1993; Vicente et al., 2007a). In our present study, the pectin
1.0 fractions changed significantly in blueberries during storage at 5 ◦
C and 10 ◦ C. The amount of WSP increased gradually with
storage time paralleled by a decreasing amount of SSP (Fig. 2A and
B). Mean- while, the amount of CSP increased during the first 21
0.5
days, and then decreased afterwards (Fig. 2C). These changes
suggested that, similar to the other fruit, pectins, as indicated by
uronic acid con- tent, were solubilized from the wall during
0.0 softening in blueberries, indicating the possible alterations in the
0 7 14 21 28 35 42 49 cross-linking of carbohy- drates in the cell walls (Manning,
1993). Moreover, it should be noted that the increase in fruit
Storage time firmness in blueberries during first
(day) 7 days of storage observed in our present study might be resulted
from the CSP accumulation (Fig. 2C) in this fruit regardless of the
Fig. 1. Changes on firmness in blueberry fruit during storage at 10 ◦ C and 5 ◦ C. Val-
ues are the means ± SE. Vertical bars represent the standard errors of the means.
storage temperatures.
Asterisks indicate significant differences between the fruit stored at 10 ◦ C and 5 ◦ Additionally, in our present study, we also observed that blue-
C (Duncan’s multiple range test, * P < 0.05). berries stored at 5 ◦ C experienced higher levels of SSP and CSP
but lower WSP content as compared with those at 10 ◦ C, which
paid to the changes in pectin, hemicellulose and cellulose con- were 31.2% and 116.7% higher in levels of SSP and CSP, respec-
tent of blueberries after harvest. In this study, we examined the tively, and 14.6% lower in WSP content than those in fruit stored
possible role that cell wall modification might play in response at 10 ◦ C at the end of storage. Thus, results here suggested that
to fruit softening during storage. Cell walls were extracted to give low temperature storage somehow prevented solubilization of
these
0.6 2.8
A * * * B
* * * * * 2.1
0.4
* *
1.4
o
0.2 o 5 C
5 C
o
o
10 C 0.7
10 C
0.0 0.0
1.8 1.5
C D
Hemicellulose (%)
* *
1.2 * 1.0
* * * *
0.6 * 0.5
0.0 0.0
3.5 E
3.0
*
*
Cellulose (%)
*
2.5 *
2.0
1.5
1.0
0 7 14 21 28 35 42 49
Storage time
(day)
Fig. 2. Changes in water-soluble pectin (A), sodium carbonate-soluble pectin (B), chelator soluble pectin (C), hemicellulose (D) and cellulose (E) contents of blueberry fruit
during storage at 10 ◦ C and 5 ◦ C. Values are the means ± SE. Vertical bars represent the standard errors of the means. Asterisks indicate significant differences between
H. Chen et al. / Scientia Horticulturae 188 (2015) 44–48 51
the fruit stored at 10 ◦ C and 5 ◦ C (Duncan’s multiple range test, * P < 0.05).
10000 15000
A * * B
8000 * 12000
Polygalacturonase
Cellulase
6000 9000
* * *
4000 6000
2000 o
5 C
o
5 C 3000
o o
10 C 10 C
0 0
800 1000
C
* D
* 800
* * *
(μmol h g FW)
600 *
α-Mannosidase
*
(μmol h g FW)
β-Galactosidase
* * * 600
-1
-1
-1 -1
400
400
200
200
0 0
Storage time
(day)
Fig. 3. Changes in activities of polygalacturonase (A), cellulase (B), -galactosidase (C) and -mannosidase (D) of blueberry fruit during storage at 10 ◦ C and 5 ◦ C. Values
are the means ± SE. Vertical bars represent the standard errors of the means. Asterisks indicate significant differences between the fruit stored at 10 ◦ C and 5 ◦ C (Duncan’s
multiple range test, * P < 0.05).
polymers. It was reported that the architectural strength of cell wall-localized enzymes acting on specific, potentially highly local-
wall can be increased by the cross-linking of pectin and the ized substrates (Brummell et al., 2004). PG hydrolyzes pectin acid
associ- ation between pectin and extensin proteins (Vicente et al., along with the main chain of polygalacturonic acid, causing pectin
2007a). As higher amount of ionically and covalently bound degradation, cell wall dissolution, and ultimately, fruit softening
pectins (CSP and SSP) was maintained in blueberries stored at 5 ◦ (Wei et al., 2010). Cellulase degrades both cellulose and -1,4-
C, it probably cross-linked to other cell wall polymers and glucan backbone of xyloglucan, a hemicellulosic polysaccharide
strengthened the cell wall, which was associated with the greater abundant in cell walls of dicotyledons (Vicente et al., 2007a; Bu
firmness. et al., 2013). -Galactosidase has been characterized in association
Increasing evidence highlighted solubilization and with the removal of galactosyl residues from cell wall polymers
depolymer- ization of hemicellulose and cellulose also played during fruit softening (Wei et al., 2010). The removal of pec-
important roles in fruit softening (Wakabayashi et al., 2000). tic galactan side-chains was an important factor in the cell wall
Vicente et al. (2007b) demonstrated hemicellulose degradation changes leading to ripening-related firmness loss (Payasi et al.,
was among the main modifications of cell wall disassembly in 2009). Glycosidases such as -mannosidase and -galactosidase
blueberry fruit dur- ing development. Results found in the are a class of carbohydrate hydrolyses that act on short chain
present work show that contents of hemicellulose and cellulose oligosaccharides, which may be present in glycoproteins, and on
decreased gradually in blueberries during storage regardless of carbohydrate heteromers or homomers (Vicente et al., 2007a). In
the storage tempera- tures, lower temperatures maintained order to reveal the mechanism involved in fruit softening of blue-
higher levels of these two components in blueberries, which berries during storage, the activities of four cell wall degrading
were 50.0% and 66.7% higher, respectively, than those in fruit at enzymes such as PG, cellulase, -galactosidase and
10 ◦ C after 49 days of storage (Fig. 2D and E). It was -mannosidase were measured. Regardless of storage
documented that the rigidity of cellular walls was probably temperatures, activities of all these four enzymes in blueberries
attributed to the cellulose-hemicellulose net- work structure, showed similar changes dur- ing storage, which increased and
formed by hydrogen-bond and crosslinks between cellulose peaked firstly and then declined
microfibrils (Wakabayashi et al., 2000). The breakdown of afterwards (Fig. 3). Low temperature at 5 ◦ C remarkably
hemicellulose was revealed to result in the disassembly of the suppressed
cellulose-hemicellulose network and fruit softening (Cheng et al., activities of the four enzymes and maintained lower WSP con-
2009). On the other hand, due to the crystalline nature of tent and higher contents of hemicellulose and cellulose
cellulose, its change was related to the resistance of cell wall to compared with the fruit stored at 10 ◦ C, which suggested that
enzymatic degradation (Zhou et al., 2011). Taken together, our cold stor- age inhibited cell wall degradation by inhibiting its
results suggest that the continuous degradation of hemicellulose degrading enzymes. At the end of the storage, activities of
and cellulose is of importance to fruit softening of blueberries cellulase, - galactosidase and -mannosidase in fruit at 5 ◦ C
during storage as well as an array of other fruit (Wakabayashi et al., were 3.2-, 1.5- and 1.2-fold, higher than those at 10 ◦ C,
2000; Zhou et al., 2011; Wang et al., 2015). Cold storage respectively. Vicente et al. (2005) also found that softening
suppressed the depolymerization and degradation of cellulose process was delayed via the inhibition of cell wall degrading
and hemicellulose, thus delaying fruit softening. enzymes including PG and - galactosidase in heat treated
strawberries, which confirmed that cell wall degrading enzymes
play an important role in fruit soften- ing. Therefore, our results
3.3. Changes in cell wall degrading enzymes
indicated that the effect of cold storage on delaying softening in
blueberries fruit may be partially due to relatively lower
It was increasingly apparent that softening and textural changes
activities of PG, cellulase, -galactosidase and - mannosidase.
were brought about by the actions of a multitude of cell
4. Conclusion L.) fruit. Postharvest Biol. Technol. Deng, Y., Wu, Y., Li, Y., 2005. Changes Vicente, A.R., Ortugno, C., Rosli, H.,
86, 337–345. in firmness, cell wall Powell, A.L.T., Greve, L.C., Labavitch,
Bunea, A., Rugina˘ , D., Scont¸ a, Z., composition and cell wall J.M., 2007b.
In summary, the present Pop, R.M., Pintea, A., Socaciu, hydrolases of grapes stored in Temporal sequence of cell wall
work showed that during C., Ta˘ ba˘ ran, F., Grootaert, C., high oxygen atmospheres. Food disassembly events in developing
Struijs, K., VanCamp, J., 2013. Res. Int. 38, fruits. 2. Anal- ysis of blueberry
fruit soften- ing in Anthocyanin determination in (Vaccinium species). J. Agric. Food
769–776.
blueberries, the decline of blue- berry extracts from various Duan, J., Wu, R., Strik, B.C., Zhao, Y., Chem. 55, 4125–4130.
fruit firmness was associated cultivars and their 2011. Effect of edible coatings on Wakabayashi, K., Chun, J.P., Huber,
antiproliferative and apoptotic the quality of fresh blueberries D.J., 2000. Extensive
with increased WSP content properties in B16-F10 metastatic solubilization and depoly-
(Duke and Elliott) under
and decreased levels of SSP, murine melanoma cells. commercial storage conditions. merization of cell wall
hemicellulose and cellulose. Phytochemistry 95, Postharvest Biol. Technol. 59, 71– polysaccharides during avocado
436–444. (Persea americana) ripen- ing
Compared with those at 10 ◦ 79.
involves concerted action of
Cantína, C.M., Minas, I.S., Goulas, V., Faria, A., Oliveira, J., Neves, P.Ä.,
C, fruit stored at 5 ◦ C showed Jiménez, M., Manganaris, G.A., Gameiro, P., Santos-Buelga, C., de polygalacturonase and
greater firmness and the Michailides, T.J., Crisosto, C.H., Freitas, V., Mateus, N., 2005. pectinmethylesterase. Physiol.
2012. Sulfur dioxide fumigation Antioxidant properties of Plant. 108, 345–352.
process of softening was alone or in combination with Wang, L., Jin, P., Wang, J., Jiang, L.L.,
prepared blueberry (Vaccinium
delayed due to lower WSP CO2 -enriched atmosphere myrtillus) extracts. J. Agric. Food Shan, T.M., Zheng, Y.H., 2015.
content and higher levels of extends the market life of Effect of - aminobutyric acid on
Chem. 53, 6896–6902.
cell wall modification and
SSP, hemicellulose and highbush blueberry fruit. Giongo, L., Poncetta, P., Loretti, P.,
senescence in sweet cherry
Postharvest Biol. Technol. 67, 84– Costa, F., 2013. Texture profiling
cellulose. Meanwhile, cold 91. of blueberries (Vaccinium spp.)
during storage at 20 ◦
C. Food
Chem. 175, 471–477.
storage also inhibited Carrington, C.M.S., Greve, L.C., during fruit development,
Wang, S.Y., Chen, C.T., Yin, J.J., 2010.
activities of cell wall Labavitc, J.M., 1993. Cell wall ripening and storage. Postharvest
Effect of allyl isothiocyanate on
metabolism in ripening fruit. Biol. Technol. 76, 34–39.
degrading enzymes such as Plant Physiol. 103, 429–434. Koca, I., Karadeniz, B., 2009.
antioxidants and fruit decay of
blueberries. Food Chem. 120,
PG, cellulase, -galactosidase Cheng, G., Duan, X., Jiang, Y., Sun, J., Antioxidant properties of
199–204.
and -mannosidase. The Yang, S., Yang, B., He, S.G., Liang, H., blackberry and blueberry fruits
Luo, Y.B., 2009. Wei, J., Ma, F., Shi, S., Qi, X., Zhu, X.,
grown in the Black Sea Region of
combined effect of cold Modification of hemicellulose Turkey. Sci. Hortic. 121, 447–450.
Yuan, J., 2010. Changes and
postharvest regulation of activity
storage on these enzymes polysaccharides during ripening Li, X., Nakagawa, N., Nevins, D.J.,
of postharvest banana fruit. Food and gene expression of enzymes
could slow down pectin Sakurai, N., 2006. Changes in the
related to cell wall degradation
Chem. 115, 43–47. cell-wall polysac- charides of
solubilization and Connor, A.M., Luby, J.J., Hancock, J.F., outer pericarp tissues of kiwifruit
in ripening apple fruit.
depolymeriza- tion which Berkheimer, S., Hanson, E.J., 2002. Postharvest Biol. Technol. 56,
during development. Plant
Changes in fruit antioxidant 147–154.
was related to the firmer Physiol. Biochem. 44, 115–124.
Zheng, Y., Wang, C.Y., Wang, S.Y.,
activity among blueberry Manning, K., 1993. Soft fruit. In:
structure in blueberries cultivars during cold- Zheng, W., 2003. Effect of high-
Seymour, G.B., Taylor, J.E., Tucker,
stored at 5 ◦ C. temperature storage. J. Agric. G.A. (Eds.), Bio- chemistry of Fruit
oxygen atmo- spheres on
Food Chem. 50, 893–898. blueberry phenolics,
Ripening. Chapman and Hall,
anthocyanins, and antioxidant
Cambridge, UK, pp. 347–377.
Acknowledgements Pathak, N., Sanwal, G.G., 1998.
capacity. J. Agric. Food Chem. 51,
7162–7169.
Multiple forms of
Zhou, R., Li, Y.F., Yan, L.P., Xie, J.,
This study was supported polygalacturonase from banana
2011. Effect of edible coatings on
fruits. Phytochem 48, 249–255.
by the National Natural enzymes, cell membrane
Paul, K.K., Jerome, P.V.B., 1982.
Science Foun- dation of integrity, and cell-wall
Carbohydrate interference and its
constituents in relation to
China (31471635) and correction in pectin analysis
brittleness and firmness of
using the m-hydroxydiphenyl
Special Fund for Agro- Huanghua pears (Pyrus pyrifolia
method. J. Food Sci. 47, 756–759.
scientific Research in the Nakai, cv. Huanghua) during
Payasi, A., Mishra, N.N., Chaves, A.L.S.,
storage. Food Chem. 124, 569–
Public Interest (201303073). Singh, R., 2009. Biochemistry of fruit
575.
softening:
an overview. Physiol. Mol. Biol.
References Plants 15, 103–113.
Sethu, K.M.P., Prabha, T.N.,
Tharanatha, R.N., 1996. Post-
Angeletti, P., Castagnasso, H., Miceli,
harvest biochemical changes
E., Terminiello, L., Concellón, A.,
associated with the softening
Chaves, A., Vicente, A.R., 2010.
phenomenon in Capsicum
Effect of preharvest calcium
annuum fruits. Phyto- chemistry
applications on postharvest
42, 961–966.
quality, softening and cell wall
Smith, M.A.L., Marley, K.A., Seigler, D.,
degradation of two blueberry
Singletary, K.W., Meline, B., 2000.
(Vaccinium corym- bosum)
Bioactive properties of wild
varieties. Postharvest Biol.
blueberry fruits. J. Food Sci. 65,
Technol. 58, 98–103.
˙ 352–356.
Bingül, I ., Bas¸ aran-Küc¸ ükgergin,
C., Tekkes¸ in, M.S., Olgac¸ , V., Dog˘ Torri, E., Lemos, M., Caliari, V.,
ru-Abbasog˘ lu, Kassuya, C.A.L., Bastos, J.K.,
S., Uysal, M., 2013. Effect of Andrade, S.F., 2007. Anti-
blueberry pretreatment on inflammatory and
diethylnitrosamine- induced antinociceptive properties of
oxidative stress and liver injury blueberry extract (Vaccinium
in rats. Environ. Toxicol. corymbosum). J. Pharm.
Pharmacol. Pharmacol. 59, 591–596.
36, 529–538. Vicente, A.R., Costa, M.L., Martínez,
Brummell, D.A., Dal Cin, V., Crisosto, G.A., Chaves, A.R., Civello, P.M.,
C.H., Labavitch, J.M., 2004. Cell 2005. Effect of heat treatments
wall metabolism during on cell wall degradation and
maturation, ripening and softening in strawberry fruit.
senescence of peach fruit. J. Postharvest Biol. Technol. 38,
Exp. Bot. 55, 213–222.
2029–2039. Vicente, A.R., Saladié, M., Rose, J.K.,
Bu, J., Yu, Y., Aisikaer, G., Ying, T., Labavitch, J.M., 2007a. The
2013. Postharvest UV-C linkage between cell wall
irradiation inhibits the metabolism and fruit softening:
production of thylene and the looking to the future. J. Sci. Food
activity of cell wall-degrading Agric. 87,
enzymes during softening of 1435–1448.
tomato (Lycopersicon esculentum