Crop Protection 19 (2000) 393}399

Polar * a potent Polyoxin B compound for controlling powdery

mildews in apple and nectarine trees, and grapevines
M. Reuveni *, H. Cohen
, T. Zahavi, A. Venezian
Golan Research Institute, University of Haifa, P.O. Box 97, Qasrine 12900, Israel

Morag, P.O. Box 200, Qasrine 12900, Israel
Extension Service, Ministry of Agriculture, Zefat 13100, Israel
Tivonchem Ltd., P.O. Box 33428, Haifa 31333, Israel

Abstract

The e$cacy of Polar, a polyoxin B compound, in controlling powdery mildews was evaluated in commercial orchards and a
vineyard in 1997}1999 in the Golan region of Israel. Foliar sprays of 0.0125}0.05% (v/v) suspensions of Polar [1-+5-N-(5-O-
carbomonyl-2-amino-2-deoxy-L-xylonyl)-5-amino-5-deoxy-b-D-allofur anosyl uronic acid,-5-hydroxymethyluracil], sulfur, the new
strobilurin fungicide Kresoxim-methyl, and sterol inhibitor (SI) fungicides inhibited the development of the powdery mildew fungi on
leaves, fruits and leaves, and fruit clusters of "eld-grown apple trees, nectarine trees, and grapevines, respectively. The e!ectiveness of
polyoxin B in controlling powdery mildews on apple trees and grapevines was similar to that of the standard treatment with the SI
fungicides or sulfur. Applications of polyoxin B on nectarine trees provided better control of Sphaerotheca pannosa (Wallr.) Lev. than
did the SI fungicides. Alternating treatment of polyoxin B with the SI fungicide, myclobutanil enhanced the inhibitory e!ect against
Podosphaera leucotricha on apples and provided the best control of powdery mildew. These results indicate that the use of polyoxin
B enables the number of SI fungicide treatments against powdery mildew to be reduced by up to 50%. Application of polyoxin B was
not phytotoxic to plant tissue, and was more e!ective against mycelial growth and sporulation of powdery mildew fungi than against
germination of conidia. At concentrations of 1, 10 and 100 mg/l polyoxin B inhibited germination of Uncinula necator conidia, by
52.5, 76.6 and 100%, respectively. A foliar spray (100 mg/l) suspension of Polar on greenhouse-grown grapevine leaves before
inoculation with U. necator provided '90% protection against powdery mildew development, while spraying on mildewed leaves
bearing sporulating colonies of U. necator suppressed the fungus. The inhibitory e!ectiveness of polyoxin B makes it a good candidate
for the control of powdery mildews on grapevines, apple, and especially on nectarine trees.  2000 Elsevier Science Ltd. All rights
reserved.

Keywords: Disease control; Podosphaera leucotricha; Sphaerotheca pannosa; Uncinula necator

1. Introduction may cause considerable crop losses, which may reach

Powdery mildew is widely distributed and is a major
foliar disease of "eld-grown grapevines, apple and nectar-
ine trees. In grapevines, cluster and blossom infection
with Uncinula necator (Schw.) Burr. before, or shortly
after bloom may result in poor fruit set, considerable
crop loss and a decrease in wine quality (Pearson and
Goheen, 1988). Similarly, attacks of Sphaerotheca pan-
nosa (Wallr.) Lev.on young nectarine fruits and leaves
* Corresponding author. Tel.: 972-6-6962401; fax: 972-6-6961930.
E-mail address: mreuveni@research.haifa.ac.il (M. Reuveni).
50% (Grove, 1995). The fungus can infect young fruits in
early spring, causing necrotic lesions that lead to surface
cracking (Grove, 1995). Apple leaves, blossoms, buds and
fruit can be infected by Podosphaera leucotricha (Ell.
and Ev.) E. S. Salmon, which can cause russeting of fruit
and considerable crop loss (Hickey and Yoder, 1990).
Reduction of primary inoculum and timely application of
e!ective fungicides, such as sulfur, benzimidazoles and
sterol biosynthesis inhibitors, are the principal methods
of disease control in the "eld.
The intensive use of fungicides on grape in Europe and
the USA has resulted in increased frequency of strains of
powdery mildew pathogen with reduced sensitivity to
benzimidazoles and sterol inhibitors (Steva and Clerjeau,

0261-2194/00/$ - see front matter  2000 Elsevier Science Ltd. All rights reserved.
PII: S 0 2 6 1 - 2 1 9 4 ( 0 0 ) 0 0 0 3 0 - 2
394 M. Reuveni et al. / Crop Protection 19 (2000) 393}399
1990; Ypema et al., 1997). Most resistant strains survive
for several years; therefore, further applications of fungi-
cides from the same group increase the risk of reinforcing
resistant populations of the pathogen (Dekker, 1987).
In Israel, because of mild winter temperatures and
highly favorable environmental conditions during the
spring, eight or more applications of various sterol in-
hibitor fungicides are needed during the growing season
to achieve su$cient powdery mildew control on apple. In
order to reduce the intensive use of the sterol inhibitor
fungicides, alternative compounds with a di!erent mode
of action are required for disease control.
Foliar applications of inorganic salts such as sodium
bicarbonate (Homma et al., 1981; Horst et al., 1992), or
potassium silicate (Bowen et al., 1992), mineral or plant
oils (Horst et al., 1992; Northover and Schneider, 1996),
or mono-potassium phosphate fertilizer (Reuveni and
Reuveni, 1995, 1998) have been shown to reduce the
severity of powdery mildew.
Polyoxins, microbial origin fungicides, were dis-
covered in 1963 in cultures of Streptomyces cacaoi var.
asoensis in Japan (Isono et al., 1965). Among the various
components of polyoxins, polyoxin B shows the highest
antimicrobial activity and is practically used for control
of various fungal diseases of vegetables, #owers, tobacco
plants and others (Isono et al., 1965; Hwang and Yun,
1986). These compounds have already been registered in
Israel for use against powdery mildews on ornamental
and vegetable crops, but their e$cacy has yet not been
tested on grapevines and deciduous trees. We present
here data on the control of powdery mildews in apple
and nectarine trees, and grapevines by polyoxin B in a
major growing area in Israel.
2. Materials and methods
2.1. Experimental fungicides
Polar, a polyoxin B compound +1-+5-N-(5-O-carbo-
monyl-2-amino-2-deoxy-L-xylonyl)-5-amino-5-deoxy-b-
D-allofuranosyl uronic acid,-5-hydroxymethyluracil,
50% WG (Wettable Granular), Kaken Pharmaceutical,
Ltd, Japan) was used in all experiments. The concentra-
tions of the growth chambers experiments are presented
as mg a.i./l.
The sterol inhibitor (SI) fungicides myclobutanil (Sis-
tan (Nova) 125 EC, Rohm and Haas, USA), penconazole
(Topas, 100 EC Novartis, Basel Switzerland), pyrifenox
(Dorado, 480 EC, Novartis), hexaconazole (Anvil S, 50
EC Zeneca UK), were used as standard treatments in
"eld experiments. In addition, the strobilurin kresoxim-
methyl (Strobi, 50 WG, BASF AG, Germany) was
included in experiments on apple and nectarine trees.
Wettable sulfur (90 WP, Machteshim-Agan, Israel) was
also included as a standard.
2.2. Growth chambers experiments
2.2.1. Germination of U. necator and preventative activity
of Polar on grape plants
Plants. Grapevine seedlings (Vitis vinifera L.) of
`Emerald Rieslinga were grown in 10-cm diameter pots
containing a peat, perlite and soil (2 : 1 : 1 v/v/v) mixture,
in a growth chamber at 253C, and light intensity of
100}120 lE m\ s\ with a 16 h photoperiod. Plants
were fertilized twice weekly with a 0.1% 20}20}20
(N}P}K) fertilizer solution until they produced 6}8
leaves.
2.2.2. Pathogen and inoculation
An isolate of U. necator, obtained from an infected
vineyard in the Golan region, Israel, was maintained on
reinfected vines in a growth chamber (233C, 100}120 lE
m\ s\, 16 h photoperiod). Inoculum was obtained
from freshly sporulating colonies, 8}12 days after inocu-
lation. Plants were inoculated with conidia of U. necator,
gently shaken from powdery-mildewed plants.
2.2.3. Germination test
For germination tests, conidia were shaken onto glass
slides previously coated with 1% water agar containing
0, 1, 10, 50 and 100 mg/l of Polar (Table 1). Slides were
placed in Petri dishes containing wet "lter paper and
kept in the dark at 203C for 16 h. The number of ger-
minated conidia was then counted under a microscope.
2.2.4. Preventative activity
In order to test the preventive e!ect of Polar against
U. necator, a suspension of 1, 10 or 100 mg/l of Polar or
water (Table 1) was sprayed on the upper surface of each
leaf of each plant (six plants per treatment). Twenty-four
hours after spraying, plants were inoculated with conidia
of U. necator and incubated in a growth chamber for
disease development, under similar conditions as de-
scribed above. The percentage of the leaf area a!ected by
mildew was evaluated 9 days after inoculation.
2.3. Field experiments
Field experiments using apple trees, nectarine trees
and grapevines were conducted in commercial orchards
and a vineyard in 1997}1999 in the Golan region of
Israel. All cultivars were susceptible to the tested patho-
gens, and diseases had been evident in these orchards
and vineyard in previous years. Methods of fertilization,
irrigation and other cultural practices for these crops
were as recommended to commercial growers by the
Extension Service of the Ministry of Agriculture, Israel.
The annual rainfall in this region during the winter is
750}850 mm, and the average midday relative humidity
(RH) and temperature in summer are 35}40% and 303C,
 M. Reuveni et al. / Crop Protection 19 (2000) 393}399 395

respectively. The sky is cloudless during most of the
summer. Occasionally, during some nights (from the end
of May until August), the temperature falls to 14}203C
and dew accumulates on the leaf surface.
2.4. Experimental design
Apple: Applications of fungicides on apple trees began
at the tight-cluster stage and continued until terminal
shoot growth ceased. Sprays were applied to run-o!
(2500 l/ha) with a 100-l gun-sprayer (1400 KPa), at the
time intervals speci"ed for each experiment on `calen-
dara basis as recommended in this region. Spacings be-
tween apple trees were 2;4 m with a bu!er row between
treatments. Treatments were arranged in a randomized
complete block design. Plots were four apple trees rep-
licated "ve times in all experiments.
In 1997, we compared the e$cacy of Polar with that of
sulfur. The three treatments consisted of non-treated
control, wettable sulfur (1%), and Polar (0.025% v/v),
were applied to cv. &Jonathan' (12-year old trees, 3 m in
height on MM106 rootstock), and were arranged as
described above (Table 2).
In 1998, four treatments consisting of a non-treated
control, Polar at two concentrations (0.025 and 0.05%
v/v), and penconazole (0.0125% v/v) as a standard, were
applied to cv. &Jonathan' (4-year old trees, 2 m in height
on MM106 rootstock) and arranged as described above.
Five foliar sprays of each suspension were applied at
various intervals, starting on 15 April 1998 at the tight-
cluster stage, about 2 weeks before bloom. Further ap-
plications were made on 22 and 29 April, and on 6 and 17
May 1998. Disease rating of the secondary infection,
based on the percentages of mildewed leaf area and
infected leaves was determined on 21 May 1998 and will
be described in detail below. Another experiment in 1999
included the following "ve treatments: a non-treated
control, sterol inhibitor (SI) fungicide myclobutanil
(0.05% v/v) as a standard, the new strobilurin, Kresoxim-
methyl (0.015% v/v), Polar (0.025% v/v), and a treatment
with alternating applications of Polar and myclobutanil
(Table 3). Eight foliar sprays of the fungicides were ap-
plied to cv. &Jonnee' (15-year old trees, 2.5 m in height, on
M.9 rootstock), as speci"ed in Table 3.
Nectarine: Applications of fungicides on nectarine trees
began at petal fall stage. Sprays were applied to run-o!

Table 1
Germination of conidia of U. necator on Polar-amended water agar and powdery mildew development on Polar-treated grapevine leaves

Concentration (mg/l) Germinated conidia (%) Inhibition as to control (%) Infected leaf area (%) Reduction as to
control (%)

0.0 59.0 a * 54.0 a *

1.0 28.3 b 52.5 31.0 ab 42.6
10.0 13.8 c 76.6 26.3 ab 51.3
50.0 1.0 d 98.3 n.t. *
100.0 0.0 d 100.0 4.0 b 92.3

Germination of conidia was determined on glass slides previously coated with 1% water agar containing various concentrations of Polar.
Relative to germination of conidia on controls.
The preventive activity of Polar was determined 9 days after inoculation of grapevine leaves with U. necator.
Mean values within columns followed by di!erent letters are signi"cantly (p(0.05) di!erent according to Duncan's multiple range test.
Not tested.

Table 2
E$cacy of foliar sprays of Polar and sulfur on the control of powdery mildew in "eld-grown cv. &Jonathan' apple trees in 1997

Treatment Infected shoots and bloom Leaf area infected Infected leaves Leaf area infected Infected
clusters/tree (%) (%) (%) leaves (%)

28 May 1997 3 July 1997

Control 30.8 a 46.7 a 100.0 a 41.5 a 98.0 a
Polar (0.025% v/v) 12.2 b 5.3 b 46.0 b 3.2 b 32.0 b
Sulfur (1%) 15.6 b 4.1 b 36.0 b 3.2 b 29.0 b

Eight foliar sprays of Polar (0.025% v/v), or wettable sulfur (1%) were applied during the growing season on: 16, 23 and 30 April, 8, 14 and 22 May,
and 4 and 17 June 1997, as described in Section 2.
Represents the primary infection as determined on 5 May 1997 after three foliar applications.
Represents secondary infection on leaves as evaluated on 28 May, based on the percentage of leaf area infected with powdery mildew and the
percentage of leaves infected.
Represents secondary infection on leaves as evaluated on 3 July 1997.
Means within columns followed by di!erent letters are signi"cantly (p(0.05) di!erent according to Duncan's multiple range test.
396 M. Reuveni et al. / Crop Protection 19 (2000) 393}399

Table 3 Table 4
E$cacy of foliar sprays of Polar, myclobutanil and kresoxim-methyl on E$cacy of Polar and two SI fungicides in controlling powdery mildew
control of powdery mildew in "eld-grown cv. &Jonnee' apple trees in in fruit and leaves of cv. &Fantasia' nectarine trees (Exp. 1, 1998)
1998
Treatment Infected fruit Infected Infected
Treatment Primary Infected leaf Infected area (%) fruit (%) leaves (%)
infections (%) area, (%) leaves (%)
Control 11.13 a 74.0 a 53.2 a
Control 23.3 a 46.7 a 98.4 a Myclobutanil (0.05% v/v) 2.99 b 34.0 b 12.8 b
Myclobutanil (0.05% v/v) 13.7 b 0.4 b 12.4 d Penconazole (0.0125% v/v) 2.46 b 33.8 b 11.5 b
Kresoxim-methyl 17.4 b 1.0 b 23.6 c Polar (0.025% v/v) 0.32 b 7.0 c 0.4 c
(0.015% v/v) Polar (0.05% v/v) 0.05 b 1.0 c 0.0 c
Polar, 0.025% v/v 17.5 b 4.0 b 39.6 b
Myclobutanil/Polar 12.7 b 0.3 b 9.8 d Six foliar sprays of Polar (0.025 and 0.05% v/v), penconazole
(0.0125% v/v) and myclobutanil (0.05% v/v) were applied during the
Eight foliar sprays of Polar (0.025% v/v), Kresoxim-methyl (0.015% growing season starting at petal fall, on: 8 April 1998. Further applica-
v/v), myclobutanil (0.05% v/v), or alternating treatment of Polar and tions were made on 19 and 29 April, 10 and 24 May and 4 June 1998.
myclobutanil, were applied during the growing season on: 26 March, 2, The percentages of infected fruit area and infected fruit were deter-
7, 16, 23 and 30 April, 8 and 14 May 1998. Myclobutanil (0.05% v/v) or mined on 24 May 1998.
Polar (0.025% v/v) were applied only four times each in alternation, Mean percentages of infected leaves per treatment, as determined on
starting with myclobutanil. 18 June 1998.
The percentage of primary infections per tree was determined on 12 Means within columns followed by di!erent letters are signi"cantly
April and the percentages of infected leaf area and infected leaves were (p(0.05) di!erent according to Duncan's multiple range test.
determined on 14 May, as described in Section 2.
Means within columns followed by di!erent letters are signi"cantly
(p(0.05) di!erent according to Duncan's multiple range test.
Table 5
E$cacy of Polar and myclobutanil in controlling of powdery mildew in
leaves of cv. &Fantasia' nectarine trees (Exp. 2, 1998)

(2500 l/ha) with a 100-l gun-sprayer (1400 KPa), at the Percentage of infected leaves on
time intervals speci"ed for each experiment on &calendar' Treatment 29 June 14 July 27 July
basis as recommended in this region. Spacings between
nectarine trees were 4;4.5 m with a bu!er row between Control 83.0 a 83.0 a 88.8 a
treatments. Treatments were arranged in a randomized Myclobutanil (0.05% v/v) 58.3 b 51.2 b 76.5 b
complete block design. Plots were four nectarine trees Polar#Bio-"lm 6.2 c 0.8 c 10.7 c
Polar (0.025% v/v) 4.3 c 4.3 c 12.2 c
replicated "ve times in all experiments. Polar (0.05% v/v) 1.5 c 0.5 c 11.2 c
Experiments in 1998: Five treatments comprising a
non-treated control, the SI fungicides myclobutanil Eight foliar sprays of Polar at 0.025 or 0.05% (v/v), Polar # Bio-"lm
(0.05% v/v) and penconazole (0.015% v/v) as standards, and myclobutanil (0.05% v/v) were applied during the growing season,
and two concentrations of polyoxin B (0.025 and 0.05% starting at petal fall, on 7 April 1998. Further applications were made
on 21 April, 7 and 21 May, 10, 22 and 29 June and 7 July 1998.
v/v) (Table 4), were applied to cv. &Fantasia' trees (8 years Mean numbers within columns followed by di!erent letters are signif-
old, on 198 peach rootstock) in the "rst experiment. The icantly (p(0.05) di!erent according to Duncan's multiple range test.
second experiment included a non-treated control, my-
clobutanil (0.05% v/v) as a standard, two concentrations
of polyoxin B alone (0.025 and 0.05% v/v), and polyoxin
B (0.025% v/v) together with the surfactant, Bio-"lm mined on 3 June 1999 and will be described in detail
(0.05% v/v) (Table 5), which were applied to &Fantasia' below.
trees (6 years old on 198 peach rootstock). Treatments in Grapevine: Applications of fungicides on grapevines
both experiments were arranged as described above and began at 12}14-cm shoot length and continuing until
the disease assessment procedure will be described in &veraison'. Six foliar sprays were applied to run-o!
detail below. (2000 l/ha) with a 100-l gun-sprayer (1400 KPa), at ap-
In 1999, the treatments included a non-treated control, proximately 14-day intervals as recommended in this
the SI fungicide hexaconazole (0.04% v/v) as a standard, region. Spacings between vines were 1.5;3 m with a buf-
two concentrations of Polar (0.0125 and 0.025%) and fer row between treatments. Treatments were arranged in
Kresoxim-methyl (0.015% v/v). Six foliar sprays were a randomized complete block design. Plots consisting of
applied to cv. &Autumn-free' trees (8 years old on 198 six}eight adjacent vines were replicated "ve times in both
peach rootstock), starting at petal fall on 30 March 1999. experiments. Basal leaf removal, a routine practice in this
Further applications were made on 9 and 19 April, and 2, region, was conducted in the vineyard 4 weeks after
12 and 26 May 1999. Treatments were arranged as de- bloom by removing leaves around the clusters on one
scribed above. The percentage of infected fruit was deter- side of each row.
 M. Reuveni et al. / Crop Protection 19 (2000) 393}399 397

In 1998, four treatments comprising a non-treated
control, the SI fungicides myclobutanil (0.03% v/v) alter-
nating with pyrifenox (0.01% v/v) as a standard, and two
concentrations of Polar (0.025 and 0.05% v/v) (Table 6),
were applied to cv. &Chardonnay' (11 years old on Richter
110 rootstock) grapevines. In 1999, the treatments com-
prised a non-treated control, the SI fungicides my-
clobutanil (0.03% v/v) alternating with pyrifenox (0.01%
v/v) as a standard, wettable sulfur (1%), and two concen-
trations of Polar (0.0125 and 0.025% v/v) (Table 6).
Treatments were applied to cv. &Chardonnay' (12 years
old on Richter 110 rootstock) grapevines and arranged as
in 1998.
2.5. Assessment of powdery mildew on each crop
Apple: The number of primary infections on each of the
two central apple trees in each plot was determined at the
bloom stage. This refers to infected blossoms and vegeta-
tive shoots, which have emerged mildewed in the spring
as a result of the fruit bud or terminal bud of an apple
shoot becoming infected the previous season. For the
secondary infections (mildew colonizing the newly devel-
oping shoots and leaves), 10 shoots from the upper ex-
tremities of each of the two central apple trees were
randomly selected in each plot. Each of the "ve youngest
expanded leaves of each shoot was evaluated for disease
severity, based on the percentage of the mildewed leaf
area. In addition, in order to determine disease incidence,
the percentage of infected leaves in each treatment was
determined.
Nectarine: The incidence of powdery mildew on nectar-
ine trees was determined by counting the number of
mildewed leaves per shoot and calculating the percentage
of infected leaves. The youngest "ve expanded leaves of
each of 10 shoots of each of two central trees in each
replicate plot were assessed. For the fruits, disease sever-
ity was evaluated on 20 fruits on each tested tree, based
on the percentage of their surface area infected with
powdery mildew. In order to determine incidence, the
percentage of infected fruits per treatment was deter-
mined.
Grapevine: Fruit clusters of grapevines, which were
naturally infected with powdery mildew fungus, were
rated during the growing season. Ten fruit clusters were
randomly selected from each of the four central vines in
each plot (160 clusters per treatment) and rated for dis-
ease severity on the basis of percentage of fruit cluster
area infected with powdery mildew. The percentage of
infected fruit clusters per treatment was also determined
for disease incidence.
2.6. Statistical analysis
An arc-sin transformation was performed on the data.
Analysis of variance (ANOVA) using the SAS GLM (SAS
Institute, 1992) procedure was applied to the transfor-
med data. Duncan's multiple range test was applied to
determine whether di!erences between treatments were
signi"cant.
3. Results
3.1. Germination and preventive activity
Polar at 1, 10 and 100 mg/l inhibited germination of
U. necator conidia on glass slides covered with 1%
water}agar by 52.5, 76.6 and 100%, respectively, (Table 1).
Foliar spray of 10 or 100 mg/l solution of Polar on
grapevine leaves before inoculation with U. necator pro-
vided 51.3 and 92.3% protection, respectively, against
powdery mildew development (Table 1). Spraying of
100 mg Polar/l on mildewed leaves bearing sporulating

Table 6
E$cacy of Polar, sulfur and SI fungicides on control of powdery mildew in fruit clusters of "eld-grown cv. &Chardonnay' grapevine

Exp. 1, 1998 Exp. 2, 1999

Treatment Infected area (%) Infected clusters (%) Infected area (%) Infected clusters (%)

Control 32.4 a 75.0 a 81.8 a 98.5 a

Myclobutanil/pyrifenox 0.0 b 0.0 b 1.0 b 12.5 b
Sulfur n.t n.t 0.9 b 11.5 b
Polar (0.0125%) n.t n.t 0.8 b 21.0 b
Polar (0.025% v/v) 0.4 b 4.4 b 0.5 b 18.0 b
Polar (0.05% v/v) 0.0 b 0.0 b n.t n.t

Six foliar sprays were applied at 14-day intervals, during the growing season, starting on 26 April 1998 and 30 April 1999 at 12}14 cm shoot length
and continuing until veraison. Myclobutanil (0.03% v/v) and pyrifenox (0.01% v/v) were used in alternation as a standard program.
Disease ratings based on the percentage of infected fruit cluster area or percentage of infected fruit clusters were made on July 27 1998 (Exp. 1) and
July 22 1999 (Exp. 2).
Mean numbers within columns followed by di!erent letters are signi"cantly (p(0.05) di!erent according to Duncan's multiple range test.
Not tested.
398 M. Reuveni et al. / Crop Protection 19 (2000) 393}399
colonies of U. necator suppressed the fungus, as indicated
by reduction of'90% of visible symptoms of the colo-
nies and shrinkage of conidia and hyphae (data not
shown).
3.2. Field experiments
Apple: In 1997, primary infections of powdery mildew
on newly developed shoots and blossom clusters of
apples, evaluated on 5 May after three foliar applications
(21 days after the "rst application), were signi"cantly
(p(0.05) less on Polar-(0.025% v/v) and wettable sulfur
treated, than non-treated control trees (Table 2). There
were also signi"cant di!erences among the various treat-
ments, in the severity of secondary mildew (Table 2). In
general, foliar applications of Polar were as e!ective as
the standard sulfur treatment and achieved '92% re-
duction in mildewed leaf area, relative to non-treated
control (Table 2).
In the "rst experiment in 1998, foliar applications of
Polar at both concentrations provided similar signi"cant
(p(0.05) control of secondary infections of powdery mil-
dew on apple leaves to that provided by the SI fungicide
penconazole (88}95% reduction compared with non-
treated controls). The percentages of the leaf area infected
with powdery mildew on non-treated control trees, on
trees treated with penconazole (0.015% v/v), and on
those treated with Polar at 0.025 and 0.05% (v/v) were
18.9, 0.9, 2.3 and 0.8, respectively. In the second trial,
three foliar applications of Polar and fungicides reduced
the number of primary infections on newly developed
shoots and on blossom clusters of apples, compared with
controls, as recorded 17 days after the "rst application
(Table 3). Foliar sprays of Polar at 7-day intervals inhib-
ited secondary infections of powdery mildew. However,
alternating Polar with the SI fungicide, myclobutanil
signi"cantly enhanced the inhibitory e!ect to a level
similar to that of myclobutanil and slightly better than
that of new strobilurin Kresoxim-methyl fungicide
(Table 3).
Nectarine: The inhibitory e!ect of Polar in controlling
powdery mildew on fruits and leaves of nectarine trees
was more signi"cant than that of the SI fungicides
(Tables 4 and 5). It was mainly expressed as greater
reduction in disease incidence including both the number
of infected fruits and leaves by both concentrations of
Polar than those by SI fungicides (Tables 4 and 5).
Adding the surfactant Bio-"lm to Polar did not improve
its e$cacy (Table 5). Nevertheless, Polar provided excel-
lent control and was signi"cantly more e!ective than
myclobutanil (Table 5). Similar results were obtained in
the 1999 experiment. Polar e!ectively (p(0.05) control-
led powdery mildew on fruits, even when applied at a
concentration of 0.0125% (v/v). The percentages of
infected fruits on non-treated control trees, and on trees
treated with hexaconazole (0.04% v/v), Kresoxim-methyl
(0.015% v/v), or Polar at 0.0125 and 0.025% (v/v), were
54.0, 11.0, 7.0, 4.0 and 0.0%, respectively, as determined
8 days after the last foliar application.
Grapevine: Symptoms of powdery mildew on
grapevine in both years were observed initially only on
berries of non-treated control vines. Overall, foliar ap-
plications of Polar at 0.025 and 0.05% (v/v) and of SI
fungicides e!ectively (p(0.05) inhibited disease develop-
ment in 1998 (Table 6). Although disease was more severe
in 1999, Polar at lower concentrations of 0.0125 and
0.025% (v/v) provided similar protection against pow-
dery mildew to that by sulfur or SI fungicides (Table 6).
No phytotoxicity to the foliage of the various crops
was observed as a result of foliar applications of the
polyoxin B compound.
4. Discussion
This study demonstrates that the polyoxin B com-
pound, Polar can control powdery mildew on apple and
nectarine trees, and grapevines, and con"rmed previous
"ndings on the value of this compound in disease control
in other host}pathogen systems (Isono et al., 1965;
Hwang and Yun, 1986). To the best of our knowledge,
this is the "rst report to describe the e$cacy of polyoxin
B on nectarine trees and grapevines.
Systemic fungicides were equally as or more e!ective
than Polar in controlling powdery mildew on apple, but
the number of SI applications was reduced by 50% when
Polar sprays were alternated with systemic fungicides.
Alternating sprays provided better protection than did
polar alone, which could, perhaps, ensure greater #exibil-
ity for disease control, by a reduction in the use of SI
fungicides without compromising disease control. Polar
was as e!ective as the SI fungicides or sulfur in control-
ling powdery mildew on grapevines but, Polar was more
e!ective than SI fungicides against S. pannosa on nectar-
ines. Overall, observations during the 1998 season on
apple and nectarine trees, and grapevines showed that
increasing the concentration of polar from 0.025 to
0.05% (v/v) did not improve its e$cacy against powdery
mildew, but caused no phytotoxicity on plant foliage.
Moreover, the 1999 results revealed that the use of re-
duced rates of Polar (0.0125% v/v) was as e!ective as the
standard treatments for controlling powdery mildew in
nectarine trees, and particularly, in grapevines, on which
the disease was very severe in that year. This indicates
that Polar is a potent compound for controlling powdery
mildew on these crops.
Examination of various stages in the host}pathogen
relationship and the pathogenesis of U. necator on grape-
vines, may partially explain the mode of action of foliar-
applied Polar in controlling powdery mildew. The spore
germination-preventive action of polyoxin B is relatively
weak compared with that of stobilurin fungicides: Polar
 M. Reuveni et al. / Crop Protection 19 (2000) 393}399
at 10 or 100 mg/l inhibited germination of U. necator
conidia by 76.6 and 100%, respectively (Table 1), while
a concentration of 0.1 mg/l of Kresoxim-methyl provided
100% inhibition (M. Reuveni, unpublished, Ypema and
Gold, 1999). Spraying of 100 mg Polar/l on greenhouse-
grown grapevine leaves before inoculation with U.
necator, provided '90% protection against powdery
mildew development. Spraying Polar at the same concen-
tration on mildewed leaves, which bore sporulating colo-
nies of U. necator suppressed the fungus, as indicated by
reduction of '90% of visible symptoms of the colonies
and shrinkage of conidia and hyphae (M. Reuveni, un-
published). These "ndings are consistent with earlier
reports (Isono et al., 1965; Hwang and Yun, 1986),
indicating that preventive action of polyoxin B against
the mycelial growth and sporulation is quite powerful,
but that against spore germination is weak.
The mode of action of Polar is di!erent from that of
sterol inhibitor fungicides (Koller and Scheinp#ug, 1987)
or strobilurins (Ypema and Gold, 1999). Polyoxin B is
thought to inhibit fungal cell-wall chitin biosynthesis: a
large quantity of accumulated UDP-N-acetylglucosamine,
a chitin precursor, was observed in a series of experi-
ments (Isono et al., 1965). It is considered that such an
inhibitory action by polyoxin B involves the competitive
inhibition of chitin synthetase, caused by a structural
similarity between polyoxin B and UDP-N-acetyl-
glucosamine (Isono et al., 1965). Alternation of Polar in
spraying programs might reduce the possible develop-
ment of fungicide-resistant populations of powdery mil-
dew fungi (Staub, 1991).
The performance of Polar in controlling powdery mil-
dews indicates that it would be an attractive compound
for practical use in apple and nectarine orchards and in
the vineyards. This was particularly expressed when Po-
lar was used against S. pannosa on nectarines: it exhibited
more signi"cant and powerful protection than SI fungi-
cides. S. pannosa has become a serious problem increas-
ingly di$cult to control, in Israel in the last few years,
probably, because of its reduced sensitivity to SI fungi-
cides, as observed in the present study (Tables 4 and 5)
and in other trials (M. Reuveni, unpublished data). Cur-
rently, Polar provides the best control against powdery
mildew in nectarine trees in Israel, as presented in this
paper. These "ndings, together with the fact that the use
of sulfur might be limited in Israel, because of the warm
weather during the growing season, make Polar a good
candidate for powdery mildew control. A program,
which alternates applications of Polar with conventional
SI fungicides, the new strobilurin fungicides (Ypema and
Gold, 1999), sulfur and, phosphates (Reuveni and
Reuveni, 1995) may reduce intensive use of site-speci"c
fungicides against powdery mildews.
399
Acknowledgements
The authors thank D. Scheglov for his assistance.
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