Postharvest Biology and Technology 41 (2006) 16–21

Ethylene and fruit softening in the stony hard mutation in peach

Hiroko Hayama ∗ , Miho Tatsuki, Akiko Ito, Yoshiki Kashimura
Department of Plant, Cell & Environment, National Institute of Fruit Tree Science, NARO, 2-1 Fujimoto, Tsukuba, Ibaraki 305-8605, Japan

Received 16 November 2005; accepted 6 March 2006

Abstract

The stony hard (hd) peach is characterized by a lack of ethylene production and a firm flesh in mature fruit; exogenous ethylene induces
a loss of fruit firmness. The mutation is inherited independently of the M (melting/non-melting) trait that is controlled by a ripening-
related endopolygalacturonase (endoPG) gene. We studied the process of fruit softening and the activities of the three pectolytic enzymes:
endoPG, exopolygalacturonase (exoPG), and pectin methylesterase (PME), in the stony hard cv. Manami with and without ethylene treatment.
Exogenous ethylene rapidly reduced the flesh firmness of the stony hard fruit which neatly correlated with increases of endo- and exoPG
activity. The increased levels of endo- and exoPG activity resembled those detected in fruit of the normal cv. Akatsuki, which served as a
control. In contrast to PGs, PME activity was not affected by ethylene and did not correlate with flesh firmness. Thus, the stony hard mutation
does not seem related to fruit softening enzymes, but to the control of ethylene levels in the ripening fruit. Our results underline the crucial
role of ethylene in the induction of fruit softening in peach.
© 2006 Elsevier B.V. All rights reserved.

Keywords: Ethylene; Fruit softening; Peach; Polygalacturonase; Prunus persica

1. Introduction
Fruit ripening is a complex process that culminates in pro-
nounced changes in color, texture, sugar accumulation, and
aroma of the fruit flesh. With respect to their ripening mech-
anism, fruit are divided into two broadly defined groups:
climacteric, where ripening is accompanied by a peak in
respiration, and non-climacteric, where respiration shows no
such time-course. In the former, a sharp increase in climac-
teric ethylene production is considered to control the initia-
tion of the ripening process (Alexander and Grierson, 2002).
However, ripening processes differ greatly between species,
and ethylene-dependent as well as ethylene-independent reg-
ulatory mechanisms co-exist to coordinate the process in
climacteric and non-climacteric fruit (Lelièvre et al., 1997).
In peach (Prunus persica (L.) Batsch), the fruit texture
is usually classified into melting-flesh (MF) or non-melting-
flesh (NMF). MF fruit are the common table peaches which
lose flesh firmness rapidly during ripening, while NMF fruit
∗ Corresponding author. Tel.: +81 29 838 6502; fax: +81 29 838 6437.
E-mail address: hhiroko@affrc.go.jp (H. Hayama).
are suitable for processing as they lack the rapid loss of
firmness during late stages of ripening (Bailey and French,
1932). The MF phenotype has been reported to be caused
by a ripening-related endopolygalacturonase (endoPG; EC
3.2.1.15); in NMF cultivars, the gene appears to be non-
functional (Pressey and Avants, 1978; Callahan et al., 2004).
The MF phenotype is accompanied by a large increase in
the contents of soluble pectin and progressive pectin depoly-
merization (Pressey et al., 1971; Fishman et al., 1993).
Increased activities of cell wall-modifying enzymes have
been observed in MF peach cultivars, including exopoly-
galacturonase (exoPG; EC 3.2.1.67), pectin methylesterase
(PME; EC 3.1.1.11), endo-1,4-␤-glucanase (EC 3.2.1.4),
endo-1,4-mannanase (EC 3.2.1.78), ␣-arabinosidase (EC
3.2.1.55), and ␤-galactosidase (EC 3.2.1.23) as well as
endoPG (Brummell et al., 2004a). However, other factors
also seem to contribute to the softening process after harvest.
Furthermore, it has been suggested that an imbalance in activ-
ities of cell wall-modifying enzymes causes the mealy texture
which is induced by prolonged storage at low temperature
(Zhou et al., 2000; Brummell et al., 2004b). Generally, the
softening process resulting in a ‘melting’ texture is complex

0925-5214/$ – see front matter © 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.postharvbio.2006.03.006
 H. Hayama et al. / Postharvest Biology and Technology 41 (2006) 16–21
and precisely regulated, as the expression patterns of puta-
tively cell wall metabolism-related genes suggest (Trainotti
et al., 2003).
“Stony hard-flesh” is a flesh texture in peach that occurs
in cultivars with a reduced level of ethylene production, and
that are characterized by crispy fruit flesh and a lack of soft-
ening during fruit ripening (Hayama et al., 2000; Haji et al.,
2003, 2004). The stony hard trait is controlled by a single
recessive gene (hd) which is inherited independently of the
M (MF/NMF) trait (Yoshida, 1976; Haji et al., 2005). When
the stony hard is combined with Melting, the fruit reduce their
firmness through continuous exposure to ethylene (Hayama
et al., 2003; Haji et al., 2005). Therefore, the softening pro-
cess in stony hard peaches appears to be blocked by a lack
of ethylene, and not by mutations of cell wall-modifying
enzymes. However, the activities of these enzymes includ-
ing PGs have not been investigated in stony hard peaches so
far.
In this study, we analyzed the activities of endoPG, exoPG,
and PME in the stony hard flesh cv. Manami (hdhdM-) with
and without ethylene treatment, and in the MF cv. Akatsuki
(Hd-M-) to clarify the relationship of pectolytic activity and
the stony hard mutation.
2. Material and methods
2.1. Plant material
Fruit of peach (Prunus persica (L.) Batsch) cv. Akatsuki
cultivated at the National Institute of Fruit Tree Science,
Ibaraki, Japan, and of cv. Manami grown at the Nagano Fruit
Tree Experiment Station, Nagano, Japan, were hand-picked
at the commercial harvest stage. Both cultivars are mid-
season maturing, with cling stone, white flesh, and excellent
quality. The texture of the ‘Akatsuki’ fruit flesh is “melting,”
while that of ‘Manami’ is “stony hard”.
2.2. Fruit ripening and firmness
Fruit of ‘Manami’ and ‘Akatsuki’ were packed in 40 L
containers and stored at 25 ◦ C. The containers were venti-
lated with a continuous flow of CO2 -free, humidified air at
a flow rate of 7 L min−1 . For ethylene treatment, ethylene
was added to the air flow to a final concentration in the con-
tainer of 100 ␮L L−1 . Ethylene levels in the containers were
checked regularly with a gas chromatograph equipped with an
activated alumina column (GC-14A, Shimadzu Corporation,
Japan). Carbon dioxide levels also were monitored using a
Porapak Q column (GC-14A, Shimadzu Corporation, Japan);
concentrations were kept below 0.12% to avoid CO2 damage
to the fruit. Fruit were sampled at 1, 2, 3, and 5 days after
harvest. Ethylene-treated ‘Manami’ fruit were sampled addi-
tionally at 6 and 12 h after treatment. Fruit flesh firmness was
measured after removing a small disc of skin in the centers of
the two halves of the fruit as defined by the plane of the raphe;
17
the force required to insert a penetrometer (FT011, Fujihira
Industry Co. Ltd., Japan) with a plunger of 8 mm diameter
into the flesh was determined. Flesh tissue was diced, imme-
diately frozen in liquid N2 , and stored at −80 ◦ C until further
use.
2.3. Enzyme activities
All steps of enzyme extraction were conducted at 4 ◦ C.
The determination of PG activities was conducted according
to Zhou et al. (2000) with some modification. Frozen fruit
flesh (5 g) was blended in a homogenizer with 2 volumes of
12% polyethylene glycol 6000 and 0.2% sodium bisulfite.
The homogenate was centrifuged and the pellet was washed
twice with water containing 0.2% sodium bisulfite. The pellet
was incubated on a shaker for 2 h in 3.5 mL 50 mM Na acetate
buffer (pH 5.0) containing 0.5 M NaCl. Following centrifu-
gation, the supernatant was loaded on a desalting gel column
(Bio-Rad, P-6) equilibrated with 50 mM Na acetate buffer
(pH 5.0). The column was washed with 4.5 ml 50 mM Na
acetate buffer (pH 5.0) and fractions were collected. For the
determination of endoPG activity, the enzyme extract (3 mL)
was mixed with 2% polygalacturonic acid (PGA; Sigma) in
50 mM acetate (pH 4.3) and incubated at 30 ◦ C. The protein
content of the extract was determined by a DC protein assay
kit (Bio-Rad Laboratories, Inc.) using BSA as a standard.
EndoPG activity was determined in an Ostwald viscosime-
ter (Sansyo Co. Ltd., Japan) by the decrease of viscosity
in the assay mixture. Viscosity was measured immediately
after adding the enzyme extract and after 18 h of incubation.
One activity unit was defined as a 1% reduction in viscos-
ity over a period of 18 h mg−1 protein. To establish exoPG
activities, the enzyme extract (1.5 mL) was diluted 2:1 with
50 mM Na acetate buffer (pH 5.0) containing 2 mM CaCl2 ,
mixed with an equal volume of 0.5% PGA (Sigma) in 50 mM
Na acetate buffer (pH 4.3), and incubated at 30 ◦ C. ExoPG
activity was assayed by measuring the hydrolytic release of
reducing groups from PGA using a 2-cyanoacetamide assay
under conditions as described by Gross (1982). One unit of
activity was defined as 1 ␮g galacturonic acid released per
mg protein per hour.
PME extraction was performed according to Obenland and
Carroll (2000), and the activity of PME was assayed at pH
7.0 by the gel diffusion assay of Downie et al. (1998). The
activity was determined by comparison to a standard curve
made using orange peel PME (Sigma); one unit of activity
was defined as the activity of the orange peel PME standard
solution (1 ␮g mL−1 ).
3. Results
3.1. Fruit ripening
The flesh firmness of both ‘Manami’ and ‘Akatsuki’ fruit
at harvest was approximately 29N. ‘Manami’ fruit (stony
18 H. Hayama et al. / Postharvest Biology and Technology 41 (2006) 16–21
Fig. 1. Flesh firmness of peach fruit of cv. Manami treated with ethylene-
free air (‘Manami’) or with ethylene (E-‘Manami’), and cv. Akatsuki treated
with ethylene-free air (‘Akatsuki’). The data are means ± S.E. of more than
five replications.
hard-flesh) without ethylene treatment remained firm dur-
ing the ripening period of 5 days, while ethylene-treated fruit
began to decrease in firmness after 24 h of ethylene treatment
and rapidly softened within 2–3 days (Fig. 1). ‘Akatsuki’
fruit (melting-flesh) rapidly lost their firmness within 2 days
even in the absence of ethylene (Fig. 1). The texture of
‘Akatsuki’ fruit developed into a juicy melting flesh; that of
ethylene-treated ‘Manami’ was slightly less juicy and showed
water-core-like symptoms in the flesh after 5 days of ethylene
treatment (Fig. 2).
Fig. 2. Water-core symptoms appearing in ‘Manami’ fruit after 5 days of
ethylene treatment.
Fig. 3. EndoPG activity in peach fruit of cv. Manami treated with ethylene-
free air (‘Manami’) or with ethylene (E-‘Manami’), and cv. Akatsuki treated
with ethylene-free air (‘Akatsuki’). Data shown are means ± S.E. of three
replications.
3.2. Enzyme activities
The endoPG activity of ‘Manami’ fruit exposed to
ethylene-free air remained low for 5 days after harvest
(Fig. 3). In ethylene-treated ‘Manami’ fruit, the activity
sharply increased after 12 h of treatment and reached a level
similar to that of ‘Akatsuki’ within 2 days, before it decreased
again towards the end of treatment (Fig. 3). In ‘Akatsuki’
fruit, the endoPG activity increased rapidly after harvest and
remained high during the 5 days of the experimental period.
Fig. 4. ExoPG activity in peach fruit of cv. Manami treated with ethylene-
free air (‘Manami’) or with ethylene (E-‘Manami’), and cv. Akatsuki treated
with ethylene-free air (‘Akatsuki’). The data are means ± S.E. of three repli-
cations.
 H. Hayama et al. / Postharvest Biology and Technology 41 (2006) 16–21
Fig. 5. PME activity in peach fruit of cv. Manami treated with ethylene-
free air (‘Manami’) or with ethylene (E-‘Manami’), and cv. Akatsuki treated
with ethylene-free air (‘Akatsuki’). Data shown are means ± S.E. of three
replications.
In ‘Manami’ fruit, the level of exoPG remained low during
ripening without ethylene treatment (Fig. 4). In contrast, the
activity in ethylene-treated ‘Manami’ increased after 12 h of
treatment as fruit firmness decreased (Fig. 1). The exoPG
activity in ‘Akatsuki’ fruit increased steadily during ripening
and was similar to that of ethylene-treated ‘Manami’ fruit.
PME activities were more or less stable through the ripen-
ing period in ‘Manami’ and ‘Akatsuki’ and were not affected
by ethylene (Fig. 5). There was no correlation between the
level of PME activity and flesh firmness.
4. Discussion
Peach is a climacteric fruit which significantly increases
ethylene production during ripening (Tonutti et al., 1991).
However, stony hard cultivars such as ‘Manami’ produce lit-
tle ethylene and retain a crispy flesh texture (Haji et al., 2001).
They are ethylene-sensitive, though, as exogenous ethylene
induces a rapid loss of firmness (Haji et al., 2005). We here
confirmed these facts in ‘Manami’ fruit (Fig. 1) and found
that ‘Manami’ fruit firmness correlated well with the lev-
els of exoPG and endoPG activities (Figs. 4 and 5) under
all experimental conditions. These results demonstrated that
fruit softening-related enzymes are functional in the stony
hard cultivar ‘Manami’, and that ethylene is necessary to
induce their activities. On the other hand, NMF peach culti-
vars have been reported to contain significant levels of exoPG
(Pressey and Avants, 1978) and to produce ethylene (Brovelli
et al., 1999) during ripening. Therefore, the NMF texture
appears to be caused by a lack of endoPG activity.
The ‘Manami’ fruit remained firm for at least 12 h after the
beginning of ethylene treatment and became softer after 24 h
19
(Fig. 1). Similarly, the activities of endo- and exoPG activities
were constant during the first 12 h and increased significantly
after 24 h (Figs. 3 and 4). Thus, it takes at least 12–24 h after
ethylene application to activate softening-related enzymes
that reduce the fruit flesh firmness in peach. In tomato, trans-
genic fruit expressing an antisense 1-aminocyclopropane-1-
carboxylic acid (ACC) synthase gene that strongly blocked
ethylene production were shown to induce PG mRNA within
6 h of ethylene treatment, but significant increases in enzyme
activity were undetectable within 24 h (Sitrit and Bennett,
1998). It therefore seems that PG activity responds to ethy-
lene more rapidly in peach than in tomato.
In peach, exoPG as well as endoPG have been reported to
increase their activities during ripening (Pressey and Avants,
1973, 1978; Downs et al., 1992; Orr and Brady, 1993); in
contrast, levels of exoPG activity remain constant in ripening
tomato fruit (Pressey, 1987). Our results are consistent with
these previous findings; in addition, we demonstrated that
the increases of enzyme activities were induced by ethylene.
PME activities were not affected by ethylene, and a signif-
icant level of activity was observed in stony hard peaches
that produced low amounts of ethylene during ripening.
Brummell et al. (2004a) reported that PME activity increased
sharply at an early stage of ripening and remained high there-
after. Taken together, these results suggest that PME activity
is independent of ethylene or might be stimulated by low
levels of ethylene at early ripening stages.
In climacteric fruit, ripening events such as fruit softening,
chlorophyll degradation, coloring, and sugar accumulation
are considered to be induced by ethylene (Alexander and
Grierson, 2002). However, stony hard peach fruit undergo
normal changes in skin color and sugar composition on the
tree, even though the fruit produce little ethylene (Haji et al.,
2004). In a normal melting peach cultivar, ethylene produc-
tion was low in the early phases of ripening when the fruit
began to change skin color and sugar contents, and the climac-
teric peak occured only after the fruit had already softened
(Tonutti et al., 1996). Therefore, it appears doubtful whether
ethylene is the trigger of ripening events in peach. However,
as basal levels of ethylene can be observed at the early stage
of ripening in normal melting peach (Tonutti et al., 1997)
and even in stony hard peach (Tatsuki et al., 2006), some
early ripening events affecting the appearance and taste of
peach fruit may be activated via two possible pathways: by
an ethylene-independent pathway, that is, by a developmen-
tal regulation, or by a pathway that depends on low levels
of ethylene (Fig. 6). In the latter case, increases in the sen-
sitivity of the fruit or in endogenous ethylene concentration
may be decisive factors. On the other hand, rapid softening
occurring at late ripening stages requires significant levels of
ethylene. This rapid, ethylene-dependent softening is at least
in part correlated with the activities of endoPG, exoPG, and a
ripening-related expansin gene, PpExp3, which is inducible
by ethylene in stony hard fruit and is expressed during the
ripening period in normal fruit (Hayama et al., 2003). In
addition, treatment with exogenous ethylene induced the syn-
20 H. Hayama et al. / Postharvest Biology and Technology 41 (2006) 16–21
Fig. 6. A tentative model of the regulation of ethylene-dependent and
ethylene-independent ripening events in peach. See text for details.
thesis of volatiles in stony hard peach, though the amounts
and rates of production of these compounds have not been
quantified so far. Therefore, the induction of the synthesis of
volatiles that contribute to the fruit’s flavor or aroma may also
require significant ethylene levels in peach.
Mealy texture was observed in stony hard fruit softened by
exogenous ethylene, and water-core-like symptoms appeared
after 5 days of ethylene treatment (Fig. 2). Similar effects
were observed in the Japanese pear ‘Nijisseiki’ following
propylene treatment, which limited the degree of solubiliza-
tion and depolymerization of pectin polysaccharides in the
cell wall and led to a reduced endoPG activity compared
to the melting texture in the cultivar ‘LaFrance’ (Hiwasa
et al., 2004). Mealy texture also occurs after cold storage
in nectarine or peach fruit. Prolonged exposure to low tem-
perature causes unbalanced activities of numerous cell wall-
modifying enzymes such as PG and PME, and results in an
arrest of the breakdown of cell wall pectins (Ben-Arie and
Sonego, 1980; Obenland and Carroll, 2000; Zhou et al., 2000;
Dong et al., 2001; Brummell et al., 2004b). The stony hard
texture was not completely restored to a normal melting tex-
ture by exogenous ethylene in our experiments, suggesting
that the balance of cell wall-modifying activities during soft-
ening is complex and highly regulated. Further analysis of
stony hard peach cultivars will help to identify additional
enzymes or factors affecting the rapid loss of firmness during
fruit ripening in peach.
Acknowledgements
The authors thank Dr. Ann Callahan of the U.S. Depart-
ment of Agriculture for helpful discussion, and Mr. Katsuhiro
Tajiri of the Nagano Fruit Experiment Station, Japan, for
assistance with the sampling of the ‘Manami’ fruit.
References
Alexander, L., Grierson, D., 2002. Ethylene biosynthesis and action in
tomato: a model for climacteric fruit ripening. J. Exp. Bot. 53,
2039–2055.
Bailey, J.S., French, A.P., 1932. The inheritance of certain characters in
the peach. Proc. Am. Soc. Hortic. Sci. 29, 127–130.
Ben-Arie, R., Sonego, L., 1980. Pectolytic enzyme activity involved in
wooly breakdown of stored peaches. Phytochemistry 19, 2553–2555.
Brovelli, E.A., Brecht, J.K., Sherman, W.B., 1999. Nonmelting-flesh trait
in peaches is not related to low ethylene production rates. HortScience
34, 313–315.
Brummell, D.A., Cin, V.D., Crisosto, C.H., Labavitch, J.M., 2004a. Cell
wall metabolism during maturation, ripening and senescence of peach
fruit. J. Exp. Bot. 55, 2029–2039.
Brummell, D.A., Cin, V.D., Lurie, S., Crisosto, C.H., Labavitch, J.M.,
2004b. Cell wall metabolism during the development of chilling injury
in cold-stored peach fruit: association of mealiness with arrested dis-
assembly of cell wall pectins. J. Exp. Bot. 55, 2041–2052.
Callahan, A.M., Scorza, R., Bassett, C., Nickerson, M., Abeles, F.B.,
2004. Deletions in an endopolygalacturonase gene cluster corre-
late with non-melting flesh texture in peach. Funct. Plant Biol. 31,
159–168.
Dong, L., Zhou, H.-W., Sonego, L., Lers, A., Lurie, S., 2001. Ethy-
lene involvement in the cold storage disorder of ‘Flavortop’ nectarine.
Postharvest Biol. Technol. 23, 105–115.
Downie, B., Dirk, L.M.A., Hadfield, K.A., Wilkins, T.A., Bennett, A.B.,
Bradford, K.J., 1998. A gel diffusion assay for quantification of pectin
methylesterase activity. Anal. Biochem. 264, 149–157.
Downs, C.G., Brady, C.J., Gooley, A., 1992. Exopolygalacturonase protein
accumulates late in peach fruit ripening. Physiol. Plant 85, 133–140.
Fishman, M.L., Levaj, B., Gillespie, D., Scorza, R., 1993. Changes in
the physico-chemical properties of peach fruit pectin during on tree
ripening and storage. J. Am. Soc. Hortic. Sci. 118, 343–349.
Gross, K.C., 1982. A rapid and sensitive spectrophotometric method for
assaying polygalacturonase using 2-Cyanoacetamide. HortScience 17,
933–934.
Haji, T., Yaegaki, H., Yamaguchi, M., 2001. Changes in ethylene produc-
tion and flesh firmness of melting, nonmelting and stony hard peaches
after harvest. J. Jpn. Soc. Hortic. Sci. 70, 458–459.
Haji, T., Yaegaki, H., Yamaguchi, M., 2003. Softening of stony hard
peach by ethylene and the induction of endogeneous ethylene by 1-
aminocyclopropane-1-carboxylic acid (ACC). J. Jpn. Soc. Hortic. Sci.
72, 212–217.
Haji, T., Yaegaki, H., Yamaguchi, M., 2004. Varietal differences in the
relationship between maturation characteristics, storage life and ethy-
lene production in peach fruit. J. Jpn. Soc. Hortic. Sci. 73, 97–104.
Haji, T., Yaegaki, H., Yamaguchi, M., 2005. Inheritance and expression
of fruit texture melting, non-melting and stony hard in peach. Sci.
Hortic. 105, 241–248.
Hayama, H., Shimada, T., Haji, T., Ito, A., Kashimura, Y., Yoshioka,
H., 2000. Molecular cloning of a ripening-related expansin cDNA in
peach: evidence for no relationship between expansin accumulation
and change in fruit firmness during storage. J. Plant Physiol. 157,
567–573.
Hayama, H., Ito, A., Moriguchi, T., Kashimura, Y., 2003. Identification
of a new expansin gene closely associated with peach fruit softening.
Postharvest Biol. Technol. 29, 1–10.
Hiwasa, K., Nakano, R., Hashimoto, A., Matsuzaki, M., Murayama, H.,
Inaba, A., Kubo, Y., 2004. European, Chinese and Japanese pear fruits
exhibit differential characteristics during ripening. J. Exp. Bot. 55,
2281–2290.
 H. Hayama et al. / Postharvest Biology and Technology 41 (2006) 16–21
Lelièvre, J.-M., Latché, A., Jones, B., Bouzayen, M., Pech, J.-C., 1997.
Ethylene and fruit ripening. Physiol. Plant 101, 727–739.
Obenland, D.M., Carroll, T.R., 2000. Mealiness and pectolytic activity in
peaches and nectarines in response to heat treatment and cold storage.
J. Am. Soc. Hortic. Sci. 125, 723–728.
Orr, G., Brady, C., 1993. Relationship of endopolygalacturonase activity
to fruit softening in a freestone peach. Postharvest Biol. Technol. 3,
121–130.
Pressey, R., Hinton, D.M., Avantsm, J.K., 1971. Development of poly-
galacturonase activity and solubilization of pectin in peaches during
ripening. J. Food Sci. 36, 1070–1073.
Pressey, R., Avants, J.K., 1973. Separation and characterization of
endopolygalacturonase and exopolygalacturonase from peaches. Plant
Physiol. 52, 252–256.
Pressey, R., Avants, K., 1978. Difference in polygalacturonase composi-
tion of clingstone and freestone peaches. J. Food Sci. 43, 1415–1423.
Pressey, R., 1987. Exopolygalacturonase in tomato fruit. Phytochemistry
30, 1867–1870.
Sitrit, Y., Bennett, A., 1998. Regulation of tomato fruit polygalacturonase
mRNA accumulation by ethylene: a re-examination. Plant Physiol.
116, 1145–1150.
Tatsuki, M., Haji, T., Yamaguchi, M., 2006. The involvement
of 1-aminocyclopropane-1-carboxylic acid synthase isogene, Pp-
21
ACS1, in peach fruit softening. J. Exp. Bot. 57, 1281–
1289.
Tonutti, P., Casson, P., Ramina, A., 1991. Ethylene biosynthesis dur-
ing peach fruit development. J. Am. Soc. Hortic. Sci. 116, 274–
279.
Tonutti, P., Bonghi, C., Ramina, A., 1996. Fruit firmness and ethylene
biosynthesis in three cultivars of peach (Prunus persica L. Batsch).
J. Hortic. Sci. 71, 141–147.
Tonutti,e, P., Bonghi, C., Ruperti, B., Tornielli, G.B., Ramina, A., 1997.
Ethylene evolution and 1-aminocyclopropane-1-carboxylate oxidase
gene expression during early development and ripening of peach fruit.
J. Am. Soc. Hortic. Sci. 122, 642–647.
Trainotti, L., Zanin, D., Casadoro, G., 2003. A cell wall-oriented genomic
approach reveals a new and unexpected complexity of the softening
in peaches. J. Exp. Bot. 54, 1821–1832.
Yoshida, M., 1976. Genetic studies on the fruit quality of peach varieties
III texture and keeping quality. Bull. Fruit Tree Res. Sta. 3, 1–16 (in
Japanese with English abstract).
Zhou, H.-W., Sonego, L., Khalchitski, A., Ben-Arie, R., Lers, A., Lurie,
S., 2000. Cell wall enzymes and cell wall changes in ‘Flavortop’
nectarines: mRNA abundance, enzyme activity, and changes in pectic
and neutral polymers during ripening and in woolly fruit. J. Am. Soc.
Hortic. Sci. 125, 630–637.