Вы находитесь на странице: 1из 8

ORI GI NAL ARTI CLE

Effect of temperature and water activity on in vitro


germination of Monilinia spp.
C. Casals, I. Vin as, R. Torres, C. Griera and J. Usall
IRTA, UdL-IRTA Centre, XaRTA-Postharvest, Lleida, Catalonia, Spain
Introduction
Monilinia spp. are the most important cause of postharvest
decay in peaches [Prunus persica (L.) Batsch.] and nectar-
ines [Prunus persica var. nectarine (Ait) Maxim.] world-
wide. Brown rot of stone and pome fruit is caused by
Monilinia laxa (Aderh et Rulh) Honey, Monilinia fructigena
Honey in Whetzel and Monilinia fructicola (Wint.) Honey
(Byrde and Willetts 1977). In the European Mediterranean
areas, brown rot of stone fruit is caused by the fungi
M. laxa and M. fructigena (De Cal and Melgarejo 1999). In
contrast, M. fructicola causes brown rot in India, Japan,
Republic of Korea, Oceania and many areas of North and
South America and is included in the A2 list of quarantine
organisms for Europe [organisms present in the European
and Mediterranean organization for plant protection
region (EPPO), but contained, under ofcial control]
(Bosshard et al. 2006). Brown rot postharvest losses in the
European Mediterranean areas are typically more severe
than preharvest losses and occur during storage and trans-
port, in some cases even affecting fruit at the processing
stage (Hong et al. 1997; Larena et al. 2005). When condi-
tions are favourable for disease development, postharvest
losses are typically more severe reaching values of 8090%
in some cases (Hong et al. 1997; Hong and Michailides
1998). However, the most important cause of postharvest
diseases in peaches and nectarines in Spain is M. laxa
followed by M. fructigena (isolated from 10% to 15% of
Keywords
ecological determinants, germination
percentage, lag phase, Monilinia fructicola,
Monilinia fructigena, Monilinia laxa.
Correspondence
Josep Usall Rodie , IRTA, Centre UdL-IRTA,
XaRTA-Postharvest, 191, Rovira Roure
Avenue, 25198-Lleida, Catalonia, Spain.
E-mail: josep.usall@irta.cat
2008 2101: received 9 December 2008,
revised and accepted 6 May 2009
doi:10.1111/j.1365-2672.2009.04402.x
Abstract
Aims: This study evaluated the effect of temperature (038C) and water activ-
ity (a
w
: 087099) on the lag phase prior to germination and the percentage of
germination over time for Monilinia laxa, Monilinia fructicola and Monilinia
fructigena.
Methods and Results: More than 80% of viable conidia germinated at 25C
and 099 a
w
within 2 h for M. fructicola and M. fructigena and 4 h for M. laxa.
There was no germination at 38C, and all three Monilinia spp. germinated at
0C. At the lowest a
w
(087), none of the Monilinia spp. was able to germinate
at any of the incubation temperatures studied. Whereas at 090 a
w
, conidia
were only able to germinate at 15, 25 and 30C for the three species studied,
except for M. fructicola at 15C. In contrast, at 095, 097 and 099 a
w
, germina-
tion occurred at all studied temperatures less 38C. Generally, the lag phase
was longer at low levels of a
w
(090095), and differences were more evident as
temperatures were far from the optimum (05C).
Conclusions: Germination and lag phase period were markedly inuenced by
temperature and a
w
, and in general when conditions of temperature and a
w
were suboptimal, the lag phase was longer and the percentage of germination
was lower.
Signicance and Impact of the Study: Knowledge of the germination require-
ments of this fungus is important in order to understand their behaviour in
natural situations and to provide baseline data required for the construction of
new prediction models. Our study might be used to develop a predictive model
to understand and control the disease caused by Monilinia spp.
Journal of Applied Microbiology ISSN 1364-5072
2009 The Authors
Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754 47
fruit affected by brown rot) (Larena et al. 2005). Posthar-
vest losses on pome fruit caused by M. fructigena are also
usually low around 152% (Van Leeuwen et al. 2000).
Chemical control applied at preharvest is the main method
used to prevent fruit rot. However, preharvest controls are
often reported as not being efcient (Larena et al. 2005).
Moreover, the disease cannot be controlled by postharvest
treatments too, as no chemical is allowed on picked stone
fruit in the European Union.
Conidia are widely recognized as the most important
inoculum of brown rot fungi, and the risk of deterioration
and the methods that could be used for their control
depend on the knowledge of the ecological requirements
of these conidia and their interactions with other
micro-organisms (Lacey 1989). Thus, determining the fac-
tors that affect the survival of this fungus may help in
understanding the epidemiology of brown rot and the
development of disease management strategies (Tian and
Bertolini 1999; Hong et al. 2000; Van Leeuwen et al.
2002). Several studies have dealt with the biology of the
conidia in eld and in in vitro conditions (Tamm and
Fluckiger 1993; Xu and Robinson 2000; Xu et al. 2001).
However, for the Monilinia spp. affecting stone fruit
(M. laxa, M. fructicola and M. fructigena), no information
on the comparison of the three species is available
regarding the ecological determinants in a wide range of
conditions.
Temperature and water availability (water activity: a
w
)
are the most important abiotic parameters determining
the potential for conidia germination and the growth of
propagules on the fruit surface (Magan and Lacey 1988),
being a
w
the same value of relative humidity (RH) under
equilibrium conditions.
Although many efforts have been made to determinate
the germination conditions for Monilinia spp., the
knowledge of the effect of temperature and a
w
over a wide
range is limited. Weaver (1950) reported that M. fructicola
conidia needed between 2 and 25 h to germinate at 25C
on potato dextrose agar (PDA) medium. In other studies,
was reported the effect of temperature on the germination
of M. fructicola ascospores, where after 24 h of incubation
at 15, 20 and 25C, 100% of them had been germinated
(Hong and Michailides 1998). For M. laxa, the maximum
conidial germination occurred between 15 and 25C
exposed at 100% of RH and up to 88% was reported over
a wide temperature range (530C) (Tamm and Fluckiger
1993). Moreover, it was observed that the conidia of
M. laxa germinated even at )4C (Tian and Bertolini
1999). The maximum conidia germination rate for
M. fructigena occurred in the range 2325C in free water;
and this species was able to germinate over a wide temper-
ature range (330C). In contrast, germination observed
below 97% of RH was rare (Xu et al. 2001).
In order to plan rational disease-control managements is
necessary to study the conidial germination of the pathogen
in different environmental conditions. Therefore, the aim
of this study was to determine and compare the effect of
temperature and a
w
on germination percentages and lag
times for germination for M. laxa, M. fructigena and
M. fructicola in in vitro tests.
Material and methods
Isolates
Isolates of M. laxa (CPML1), M. fructigena (CPMG1) and
M. fructicola (CPMC1) were from the collection of the
Pathology Unit, UdL-IRTA Center of Lleida, Catalonia,
classied at Department of Plant Protection, INIA,
Madrid, Spain. They were originally isolated from
decayed fruits from commercial orchards and were main-
tained on PDA (39 g l
)1
; Biokar Diagnostics, Beauvais,
France) Petri dishes amended with acetone (1%; J.T.
Baker, Deventer, Holland) and stored at 4C in darkness.
Medium
The basic medium used was PDA at pH of 56. The a
w
of
this basal medium was 099. This a
w
was modied by the
addition of known amounts of the nonionic solute, glyc-
erol, in order to obtain a
w
levels of 097, 095, 090 and
087. The a
w
of all media was checked with a a
w
meter
(AquaLab, Pullman, WA, USA).
Inoculum preparation
Isolates were subcultured onto new PDA Petri dishes and
incubated in darkness at 25C for c. 2 weeks before fruit
inoculation. To obtain heavy sporulation of Monilinia
spp., each isolate strain was inoculated on freshly har-
vested peach or nectarine. For M. laxa and M. fructicola,
peach fruit were wounded with a scalped and a mycelial
plug cultured on PDA was inserted into each wound.
Fruit were then incubated at 25C and 85% RH in dark-
ness for 57 days for M. fructicola and 710 days for
M. laxa. Low sporulation on fresh fruit by M. fructigena
was overcome by inoculating canned peach halves with a
mycelial plug of M. fructigena that was cut from the
region of an actively growing region. The canned peach
halves were then placed into a sterile glass container and
incubated at 25C and 85% RH in darkness for 57 days.
After the incubation, conidia from sporulated fruit area
were suspended in 5 ml of sterile distilled water that was
amended with one drop of wetting agent per litre
(Tween-80), previously modied with glycerol to the
required a
w
treatment. The nal a
w
of the conidial
Effect of temperature and water activity on Monilinia spp. C. Casals et al.
48 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754
2009 The Authors
suspensions were 087, 090, 095, 097 and 099. The nal
conidial concentration was adjusted to 15 10
5
coni-
dia ml
)1
using a haemocytometer.
Germination study
To carry out the germination study, 10 ll droplets of the
conidia suspensions were placed around PDA Petri dishes
with the same a
w
. Then, Petri dishes with the same a
w
were unclosed in polyethylene bags and incubated at 0, 5,
15, 25, 30, 35 and 38C. Experiments were carried out
with three replicate Petri dishes per treatment. After each
incubation period, three agar discs (5 mm diameter)
coincided with each of the placed drops were removed
from each replicate using a cork borer. At each sampling
time, discs from the same temperature and a
w
were placed
into a sterile empty Petri dish, and conidia germination
was immediately stopped by adding 1 ml of ethanol
(EtOH) 999% onto a (60 mm diameter) lter paper
placed into the Petri dish. Then, Petri dishes were closed
and stored at 4C until microscopic examination. Fifty
single conidia per disc (150 per replicate; 450 per treat-
ment) were examined. Conidia were considered germi-
nated when the germ tube was equal to or longer than the
smallest diameter of conidia. The experiments were carried
out for a maximum of 30 days. The experiment was
repeated twice.
For the germination studies, the variable measured was
the percentage of germination at different temperatures
and a
w
over time. The lag phase period for each tem-
perature and a
w
treatment was considered to have
ended when the percentage of germinated conidia was
exceeded 10%.
Results
Effect of temperature on germination
The three species of Monilinia studied were able to
germinate over a wide temperature range (035C) at
35C
0
20
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0
20
40
60
80
100
30C
Time (h)
15C
0
20
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
5C
0
20
40
60
80
100
0 1 2 3 4 5
Time (days)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0C
0
20
40
60
80
100
0 3 6 9 12 15
Time (days)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
25C
0
20
40
60
80
100
0 12 24 36 48 60 72
0 12 24 36 48 60 72
0 12 24 36 96 144 192 240
0 12 24 96 144 192 240
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
Figure 1 Effect of temperature and water activity on germination percentage of Monilinia laxa. Water activity levels are ( ) 099; ( ) 097;
( ) 095; ( ) 090 and ( ) 087. Values are the mean of three replicates and 150 conidia per replicate. Vertical bars are the standard error.
C. Casals et al. Effect of temperature and water activity on Monilinia spp.
2009 The Authors
Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754 49
099 a
w
(Figs 13), but no germination occurred at 38C
for any of the tested isolates. The optimum temperature
for germination after 4 h of incubation was in the range
1530C for the studied species (Fig. 4). Isolates of
M. fructicola and M. fructigena reached 8595% germina-
tion after 2 h of incubation at 25C and 099 a
w
while
M. laxa needed 4 h (Figs 13).
When temperature increased to 35C at 099 a
w
, the
percentage of germinated conidia reached 76% (average
of M. laxa, M. fruticola and M. fructigena) within the rst
8 h of incubation (Figs 13).
All studied species had similar behaviour of germination
at 5C and 099 a
w
having c. 62% and 82% of germination
after 24 and 48 h, respectively (Figs 13,5). At 0C, the
percentage of germination after 24 h of incubation was
around 5% for the studied species (Figs 13). However,
after 48 h, the percentage of germination reached was 30%,
71% and 78% for M. laxa, M. fructigena and M. fructicola
respectively (Figs 13, 5).
Effect of temperature and a
w
interactions on germination
In general, when temperature varied from optimum
conditions to marginal values, a decrease in germination
percentage and an increase in the length of the lag phase
were observed for the studied species. A similar pattern
was observed when a
w
was reduced, regardless of temper-
ature (Figs 13).
The minimum a
w
at which germination begins varied
with temperature; and only at 15C was related to the
studied species (Table 1). Low levels of a
w
(087) and
high temperatures (38C) prevented conidia germination
by all three species almost until 30 days. Moreover, none
of the Monilinia spp. were able to germinate when
incubation conditions were 0, 5 and 35C and 090 a
w
.
Monilinia fructicola was the only Monilinia spp. that was
not able to germinate at 15C and 090 a
w
.
Generally, the lag phase was longer at low levels of
a
w
(090095), and differences were more evident as
15C
30C
0
20
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
35C
0
20
40
60
80
100
0 12 24 36 48 60 72
0 12 24 36 48 60 72
Time (h)
0 12 24 36 48 60 72
Time (h)
0 12 24 36 48 60 72
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
25C
0
20
40
60
80
100
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0
20
40
60
80
100
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0
20
40
60
80
100
G
e
r
m
i
n
a
t
i
o
n

(
%
)
5C
0 1 2 3 4 5
Time (days)
0
20
40
60
80
100
G
e
r
m
i
n
a
t
i
o
n

(
%
)
Time (days)
0C
0 3 6 9 12 15
Figure 2 Effect of temperature and water activity on germination percentage of Monilinia fructicola. Water activity levels are ( ) 099; ( ) 097;
( ) 095; ( ) 090 and ( ) 087. Values are the mean of three replicates and 150 conidia per replicate. Vertical bars are the standard error.
Effect of temperature and water activity on Monilinia spp. C. Casals et al.
50 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754
2009 The Authors
temperatures were far from the optimum (05C)
(Table 1). At the optimal temperature of 25C, the
duration of the lag phase was less inuenced by a
w
compared with the other temperatures studied. Thus, for
M. fructicola and M. fructigena, the duration of the lag
phase was not increased by reducing a
w
from 099 to
097 a
w
and for M. laxa was increased by only 2 h.
Moreover, further reducing the a
w
to 095, the duration
of the lag phase only increased to 4 h for M. fructicola
and M. fructigena and to 8 h for M. laxa (Table 1). In
general, the duration of the lag phase of the three
Monilinia spp. studied showed similar patterns of
response to temperature and a
w
. However, the lag phase
for M. laxa tended to be equal to or longer than the lag
phases for M. fructicola and M. fructigena.
The percentage and rate of germination tended to
decrease as a
w
decreased from 099 to 090. At 5C, the
percentage of germination after 48 h of incubation, at 099
097, 095 a
w
was 82%, 69% and 8% respectively (mean of
the three Monilinia spp.). After 72 h of incubation at 5C,
the percentage of germination at 099 a
w
had a similar
proles to that at 097 a
w
(c. 80%) for the three studied
species of Monilinia spp. In contrast, at 5C and 095 a
w
,
the percentage of germination after 72 h of incubation was
lower compared with 099 and 097 a
w
and depended
on the Monilinia spp. studied. Thus, for M. laxa and
M. fructigena, it was on average 39% while for M. fructicola
was 88%. At 0C and 099 a
w
, germination after 48 h of
incubation occurred, but it depended on the Monilinia spp.
studied. For M. fructigena and M. fructicola, the percentage
of germination was c. 75% and was nearly double that for
M. laxa (c. 30%). In contrast, after 72 h of incubation, the
percentage of germination showed similar proles for the
three species studied being around 15% and 82% testing
099 and 097 a
w
respectively.
Discussion
Prior to this study, there were only partial reports about
the germination of the three Monilinia species. However,
35C
0
20
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
30C
25C
5C
Time (days) Time (days)
0C
0 3 6 9 12 15
15C
0 12 24 36 48 60 72
0
20
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0
20
40
60
80
100
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0
20
40
60
80
100
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0
0 1 2 3 4 5
12 24 36 48 60 72
0
20
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0 12 24
20
0
40
60
80
100
Time (h)
G
e
r
m
i
n
a
t
i
o
n

(
%
)
0 12 24 48 96 144 192 240
36 48 60 72
Figure 3 Effect of temperature and water activity on germination percentage of Monilinia fructigena. Water activity levels are ( ) 099; ( ) 097;
( ) 095; ( ) 090 and ( ) 087. Values are the mean of three replicates and 150 conidia per replicate. Vertical bars are the standard error.
C. Casals et al. Effect of temperature and water activity on Monilinia spp.
2009 The Authors
Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754 51
to our knowledge, this is the rst time that the effect of
temperature and a
w
on germination has been studied for
all three species over a wide range of temperature and a
w
.
The results of this study have shown that germination
of conidia is markedly inuenced by the interaction of
temperature and a
w
. The optimum temperature for
germination of M. laxa conidia at 099 a
w
was over a
wide range (1530C), and the species was even able to
germinate at suboptimal temperatures (035C), but at
38C no germination was observed. Tian and Bertolini
(1999) also observed germination of M. laxa at )4C, but
in our work we studied temperatures only up to 0C
because this is the lowest temperature at which stone fruit
are stored after harvest. In our experiments, when tem-
peratures were far from the optimum (0, 5 and 35C),
the driest condition in which germination was observed
was 095 a
w
, while at the optimum range (15 25C) ger-
mination occurred even at 090 a
w
. The dry exposure led
to reduced germination, and in addition the germination
process was considerably delayed. Nevertheless, these
results suggest that M. laxa could have the potential to
germinate in absence of free water in the host tissues.
Experiments of Tamm and Fluckiger (1993) tested a
narrower range of RH (88100%) and also suggested
that the absence of free water is a limiting but not an
excluding factor in the infection process.
Monilinia fructicola also had a wide optimum tempera-
ture range (1530C) for germination and germinated over
all temperatures studied (035C), except at 38C. Weaver
(1950) also observed germination in the tested range from
5 to 30C. But these results showed a narrower optimum
range of temperature for germination (1525C). The effect
of temperature on the ability of M. fructicola conidia to
germinate is consistent with previous reports that showed
that temperature and duration of wetness are considered
to be the most important factors that affect infection
by M. fructicola (Luo and Michailides 2001a,b, 2003).
40
60
80
100
0
20
0 10 20 30 40
G
e
r
m
i
n
a
t
i
o
n

(
%
)
Temperature (C)
Figure 4 Effect of temperature at 099 water activity on germination
percentage of ( ) Monilinia laxa; ( ) Monilinia fructicola and ( )
Monilinia fructigena, after 4 h of inoculation. Values are the mean of
three replicates and 150 conidia per replicate. Vertical bars are the
standard error.
80
100
0
20
40
60
0 1 2 3 4 5
G
e
r
m
i
n
a
t
i
o
n

(
%
)
Temperature (C)
Figure 5 Effect of temperature at 099 water activity on germination
percentage of ( ) Monilinia laxa, ( ) Monilinia fructicola and ( )
Monilinia fructigena, after 48 h of inoculation. Values are the mean
of three replicates and 150 conidia per replicate. Vertical bars are the
standard error.
Table 1 Time (in h) before germination begins (lag phase period) at different temperatures (T
a
) and water activities (a
w
) for each isolate of
Monilinia laxa, Monilinia fructicola and Monilinia fructigena
T
a
(C) a
w
M. laxa M. fructicola M. fructigena
099 097 095 090 087 099 097 095 090 087 099 097 095 090 087
38 * * * * * * * * * * * * * * *
35 2 4 4 * * 2 4 24 * * 4 4 8 * *
30 4 4 12 120 * 2 2 4 72 * 2 4 12 48 *
25 2 4 8 48 * 2 2 4 48 * 2 2 4 72 *
15 2 8 12 264 * 4 4 8 * * 4 4 8 96 *
5 24 48 72 * * 24 48 48 * * 24 48 72 * *
0 48 120 168 * * 48 72 120 * * 24 72 168 * *
We consider germination begins if more than 10% of conidia were germinated.
*Germination was not observed after 30 days.
Effect of temperature and water activity on Monilinia spp. C. Casals et al.
52 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754
2009 The Authors
Furthermore, Biggs and Northover (1988) observed that
the optimum temperature for peach infection was 12 h of
wetness in the range 22525C, which is also within the
range presented in this work.
The optimum germination range was within 1530C
for M. fructigena. However, a narrower range of optimal
temperature (2325C) was reported by Xu et al. (2001).
That difference between studies could be because of the fact
that Xu et al. (2001) used a different model to determine
the optimum range of temperature. Our results showed
that conidia can germinate over a wide range of tempera-
tures, at least from 0 to 35C. Moreover, depending on the
temperature tested, M. fructigena can germinate from 090
to 099 a
w
. In contrast, Xu et al. (2001) determined that
conidia only germinated under near saturation humidity
(97% RH). The difference to our results could be because
of the isolate used or the solute used to reduce a
w
in the
medium: we used glycerol and they used NaCl solute.
Marin et al. (1995) reported for some species of Fusarium
spp. that, at low a
w
, growth rates were higher on
glycerol-amended medium compared with NaCl or
glucose-modied medium. Although our results showed a
clear effect of a
w
on conidia germination in in vitro studies,
Xu and Robinson (2000) suggested that the effects of the
duration of wet periods on infection were very less, unless
the conidia enter directly into fruits through unhealed
wound tissues and moisture on the wound surface was
sufcient for conidia to germinate and infect.
The results of this study showed that the maximum
germination for the Monilinia species studied was
generally lower than 100% even at optimal germination
condition. A possible explanation of this phenomenon
could be mycostasis. This has been investigated as a
mechanism by which propagules are protected from
spontaneous germination in the absence of potentially
colonisable substrata, such protection being at the
expense of debilitation (Lockwood 1988). This behaviour
seems to be an intrinsic mechanism of each species, and
no information is available about why mycostasis suscep-
tible fungi have adopted such a high-risk system, while
others have evolved dormancy activation systems to
achieve the same end (Cooke and Whipps 1993).
Our study showed that lag times were markedly inu-
enced by temperature and a
w
conditions. Similar proles
for M. fructicola, M. laxa and M. fructigena were observed
by Weaver (1950), Tamm and Fluckiger (1993) and Xu
et al. (2001) respectively. Our results, frequently showed
that reducing a
w
from 099 to 097, lag phase period did
not increase or increased less than reducing a
w
from 097
to 090.
In our report when a
w
was 099 at 25C, more than 20%
of conidia for M. laxa and 85% for M. fructicola
and M. fructigena were already germinated after 2 h of
incubation. In contrast, Xu et al. (2001) results showed
that the percentage of germination for M. fructigena was
only around 60%. Our results showed that, at 0C and
099 a
w
, no germinated conidia for all studied species were
observed before 1 day of incubation but more than 80%
of conidia were germinated after 6 days also at 097 a
w
;
this showed their potential infection in peaches at this
cold-stored conditions. Similar results were observed by
Tian and Bertolini (1999). However, testing M. laxa at 5C
and after 24 h of incubation detected a little difference in
comparison with Tian and Bertolini (1999) results. Thus,
at 5C, we observed that only around 60% of conidia were
already germinated while they found up to 90%.
In conclusion, our in vitro study has provided detailed
knowledge on the ecological requirements of these species
to germinate under appropriate temperature and a
w
conditions that could take place when conidia are on the
skin surface of stone fruit during preharvest and
postharvest (040C, 80100% for temperature and RH
respectively). Hence, these fundamental aspects of the
biology of Monilinia spp. might be used to develop a
predictive model to help us in understanding the epide-
miology of brown rot and the development of disease
management strategies.
Acknowledgements
We thank Dr P. Elmer for his helpful discussion and
critical reading of the manuscript. This study was
supported by a grant RTA2005-00077-CO2 from the
Ministry of Science and Education (Spain).
References
Biggs, A.R. and Northover, J. (1988) Inuence of temperature
and wetness duration on infection of peach and sweet
cherry fruits by Monilinia fructicola. Phytopathology 78,
13521356.
Bosshard, E., Hilber-Bodmer, M., Scharer, H.J., Bunter, M.
and Duffy, B. (2006) First report of the quarantine brown
rot pathogen Monilinia fructicola on imported stone fruits
in Switzerland. Plant Dis 90, 1554.
Byrde, R. and Willetts, H. (1977) The Brown Rot Fungi
of Fruit. Their Biology and Control. Oxford: Pergamon
Press.
Cooke, R. and Whipps, J. (1993) Ecophysiology of Fungi.
Oxford: Blackwell Scientic Publications.
De Cal, A. and Melgarejo, P. (1999) Effects of long-wave UV
light on Monilinia growth and identication of species.
Plant Dis 83, 6265.
Hong, C.X. and Michailides, T.J. (1998) Effect of temperature
on the discharge and germination of ascospores by apothe-
cia of Monilinia fructicola. Plant Dis 82, 195202.
C. Casals et al. Effect of temperature and water activity on Monilinia spp.
2009 The Authors
Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754 53
Hong, C.X., Holtz, B.A., Morgan, D.P. and Michailides, T.J.
(1997) Signicance of thinned fruit as a source of the
secondary inoculum of Monilinia fructicola in California
nectarine orchards. Plant Dis 81, 519524.
Hong, C.X., Michailides, T.J. and Holtz, B.A. (2000) Mycoora
of stone fruit mummies in California orchards. Plant Dis
84, 417422.
Lacey, J. (1989) Pre-harvest and post-harvest ecology of fungi
causing spoilage of foods and other stored products. J Appl
Bacteriol 67, S11S25.
Larena, I., Torres, R., De Cal, A., Lina n, M., Melgarejo, P.,
Domenichini, P., Bellini, A., Mandrin, J.F. et al. (2005)
Biological control of postharvest brown rot (Monilinia
spp.) of peaches by eld applications of Epicoccum nigrum.
Biol Control 32, 305310.
Lockwood, J.L. (1988) Evolution of concepts associated with
soilborne plant-pathogens. Annu Rev Phytopathol 26, 93
121.
Luo, Y. and Michailides, T.J. (2001a) Risk analysis for latent
infection of prune by Monilinia fructicola in California.
Phytopathology 91, 11971208.
Luo, Y. and Michailides, T.J. (2001b) Factors affecting latent
infection of prune fruit by Monilinia fructicola. Phytopa-
thology 91, 864872.
Luo, Y. and Michailides, T.J. (2003) Threshold conditions that
lead latent infection to prune fruit rot caused by Monilinia
fructicola. Phytopathology 93, 102111.
Magan, N. and Lacey, J. (1988) Ecological determinants of
mold growth in stored grain. Int J Food Microbiol 7, 245
256.
Marin, S., Sanchis, V. and Magan, N. (1995) Water activity,
temperature, and pH effects on growth of Fusarium moni-
liforme and Fusarium proliferatum isolates from maize.
Can J Microbiol 41, 10631070.
Tamm, L. and Fluckiger, W. (1993) Inuence of temperature
and moisture on growth, spore production, and conidial
germination of Monilinia laxa. Phytopathology 83, 1321
1326.
Tian, S.P. and Bertolini, P. (1999) Effect of temperature during
conidial formation of Monilinia laxa on conidial size,
germination and infection of stored nectarines. J Phyto-
pathol 147, 635641.
Van Leeuwen, G.C.M., Stein, A., Holb, I. and Jeger, M.J.
(2000) Yield loss in apple caused by Monilinia fructigena
(Ader. & Ruhl.) Honey, and spatio-temporal dynamics of
disease development. Eur J Plant Pathol 106, 519528.
Van Leeuwen, G.C.M., Holb, I.J. and Jeger, M.J. (2002) Factors
affecting mummication and sporulation of pome fruit
infected by Monilinia fructigena in Dutch orchards. Plant
Pathol 51, 787793.
Weaver, L.O. (1950) Effect of temperature and relative humi-
dity on occurrence of blossom blight of stone fruits. Phyto-
pathology 40, 11361153.
Xu, X.M. and Robinson, J.D. (2000) Epidemiology of brown
rot (Monilinia fructigena) on apple: infection of fruits by
conidia. Plant Pathol 49, 201206.
Xu, X.M., Guerin, L. and Robinson, J.D. (2001) Effects of
temperature and relative humidity on conidial germination
and viability, colonization and sporulation of Monilinia
fructigena. Plant Pathol 50, 561568.
Effect of temperature and water activity on Monilinia spp. C. Casals et al.
54 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 108 (2010) 4754
2009 The Authors