Disease Control and Pest Management

Control of Plant Diseases by Extracts of Inula viscosa

Wenqiao Wang, B. H. Ben-Daniel, and Yigal Cohen

First author: Faculty of Life Sciences of Bar-Ilan University, Ramat-Gan 52900, Israel, and Plant Protection Institute, Hebei Academy of
Agricultural and Forestry Sciences, Baoding 071000, China; second author: Inulex Ltd., Keramim, Mobile Post Negev, Israel; and third
author: Faculty of Life Sciences of Bar-Ilan University, Ramat-Gan 52900, Israel.
Accepted for publication 24 May 2004.

ABSTRACT

Wang, W. Q., Ben-Daniel, B. H., and Cohen, Y. 2004. Control of plant ranged from 0.68 to 1.02% and 0.65 to 1.00% (wt/vol), respectively. Dry
diseases by extracts of Inula viscosa. Phytopathology 94:1042-1047. matter content in fresh leaves, paste-extract yield in dry leaves, and dis-
ease control efficacy of paste extracts were similar in leaves of I. viscosa
Leaves of Inula viscosa were collected from the field, dried, and ex- collected during May to October, suggesting that, for practical use,
tracted with a mixture of acetone and n-hexane. The oily, water-insoluble harvests can be conducted during most of the growing season. The results
pastes obtained after evaporation of the solvents were used for the control show that I. viscosa may be used as an herbal source for fungicidal
of foliar diseases in growth chambers. The pastes, either dissolved in preparations against foliar diseases caused by pathogens belonging to the
acetone or emulsified in water, effectively controlled downy mildew of families Oomycetes, Ascomycetes, and Basidiomycetes.
cucumber, late blight of potato or tomato, powdery mildew of wheat, and
rust of sunflower. Mean effective dose (concentration) required for 90% Additional keywords: Blumeria graminis f. sp. tritici, herbal extracts,
inhibition of disease values for acetone solutions and water emulsions Phytophthora infestans, Pseudoperonospora cubensis, Puccinia helianthi.

Inula viscosa (L.) Aiton (syn. Cupularia viscosa G. et G., Keramim (2 acres, Loess soil) in March 2001 and at Bar-Ilan
Dittrichia viscosa Greuter) (Compositae) (common name “sticky Farm (four rows, each 50 m long, sandy-loam soil) in March
fleabane”) is a perennial weed, native to the Mediterranean Basin. 2002. Planting was done with a distance of 0.2 m between plants
It grows on hillslopes, damp habitats, and roadsides. In folklore and 1 m between rows. During the dry season, plants were irri-
medicine, this plant is used for therapeutic purposes, such as a gated by drip irrigation. Water was supplemented with of N-P-K
diuretic, topical anti-inflammatic, and haemostatic (10). Aqueous fertilizer (20:20:20) at 5 kg/ha once every 2 weeks.
extracts of I. viscosa were shown to exhibit antifungal activity in To save labor and solvents, I. viscosa leaves should first be
vitro (12,18), and organic solvent extracts were shown to be anti- dried before extraction. For this purpose, we determined the con-
bacterial (7). Cohen et al. (4) provided evidence for the antifungal tent of dry matter in fresh leaves. At Keramim, 10 kg of leaves
activity in planta of extracts made with organic solvents, includ- were collected in April, June, and September 2002, air dried for
ing methanol, ethanol, ethylacetate, acetone, chloroform, and n- 1 week, and stored in paper bags at room temperature until used.
hexane. Using thin-layer chromatography overlay assays, seven At Bar-Ilan, leaves were collected at biweekly intervals from 10
inhibitory zones against Cladosporium cucumerinum were ob- to 15 plants in each row, with each sample composed of ≈0.25 kg
served in the extracts (4). In a recent study (21), it was found that of leaves, for a total of 1 kg/harvest. Consecutive harvests each
leaf extracts of I. viscosa were highly effective in controlling were done from another set of plants. Harvests were done during
downy mildew of grapevine, caused by Plasmopara viticola. In 1 April to 15 October 2003. Leaves from each harvest were
the present study, we were interested in following the efficacy of pooled, weighed, dried in an oven at 60°C for 24 h, weighed
extracts obtained from I. viscosa during the active growing period again to determine dry matter content, and kept in plastic bags
in the field in controlling downy mildew of cucumber, late blight until used for extraction.
of potato or tomato, powdery mildew of wheat, and rust of sun- Extraction and formulation. For each harvest, a 100-g sample
flower. Another goal of this study was to compare the efficacy of of dry leaves was extracted at 35°C for 3 h by shaking in a
acetone solutions made with the dry pastes and pastes emulsified mixture of acetone and n-hexane (9:1, vol/vol) at a ratio of 1:10
in water. (wt/vol, dry leaves/solvent). The extract was filtered through
Whatman No. 1 filter paper and vacuum dried at 45 to 50°C to
MATERIALS AND METHODS obtain an oily, dry, green paste. The paste was weighed to deter-
mine the percentage of paste in the dry leaves.
Growing of I. viscosa and collection of leaves. I. viscosa seed Results obtained from the two harvests made at Bar-Ilan during
(collected in situ at Tirat-Yehuda, 20 km east of Tel-Aviv) were each month of 2003 were used to obtain the mean (and standard
sown in Speedling trays (Hishshtil, Petach-Tiqwa, Israel) con- deviation) of dry matter content in fresh leaves and paste yield in
taining a mixture of peat and perlite (1:1, vol/vol), and grown in dry leaves, by using Super Anova (Abacus Concepts Inc.,
the greenhouse. When they reached the four- to six-leaf stage Berkeley, CA).
(≈2 months after sowing), plants were transplanted in the field at Pastes freely dissolved in acetone, producing greenish, trans-
parent solutions. Such solutions were applied as a spray to the
foliage of potted plants to assess their efficacy in controlling dis-
Corresponding author: Y. Cohen; E-mail address: [email protected] ease. Acetone solutions were prepared by dissolving 2 g of each
paste in 100 ml of acetone and diluting with acetone into a series
Publication no. P-2004-0809-01R of concentrations (2, 1, 0.5, and 0.25%, wt/vol). A water emulsion
© 2004 The American Phytopathological Society was made by melting 34.4 g of a paste (derived similarly from

1042 PHYTOPATHOLOGY
leaves of I. viscosa collected at Keramim in June 2002) in a water calculated similarly. Analysis of variance was performed followed
bath (50°C), adding 65.6 g of emulsifier into the paste, and vor- by Fisher’s protected least significant difference (LSD) test to
texing the mixture for 2 min to form a 34.4% emulsified concen- establish significant differences (P = 0.05) in ED90 values among
trate (EC) (for details, see patent application IL149,716, May pathosystems.
2002) This product formed a stable emulsion when mixed with Efficacy of extracts emulsified in water. Paste derived from
water. leaves of I. viscosa collected in June 2002 was formulated in June
Plants and pathogens. Cucumber (Cucumis sativum L.) 2003 into a 34.4% EC product. Formulated paste and blank con-
‘Nadyojni’, susceptible to Pseudoperonospora cubensis (Berk. & trols (containing the same rate of emulsifier only) were diluted
Curt. et de Toni) Rost.; potato (Solanum tuberosum L.) ‘Mondial’ with water into emulsions containing 1, 0.5, 0.25, or 0.125%
and tomato (Lycopersicon esculentum Mill.) ‘ZH’, both suscep- paste (wt/vol). Emulsions were sprayed onto the foliage of plants,
tible to Phytophthora infestans Mont. (de Bary); wheat (Triticum with three replicate plants (or pots for wheat) per concentration.
sativum L.) ‘Sharon’, susceptible to Blumeria graminis f. sp. Plants treated with water only were used as an untreated control.
tritici (DC.) Speer; and sunflower (Helianthus annuus L.) ‘H- Treated plants, blank-control plants, and water-treated controls
567’, susceptible to Puccinia helianthi Schwein. were grown then were inoculated with their respective pathogens and incu-
from seed (tubers for potato) in 1-liter pots containing peat + per- bated as mentioned above. The percentage of infected leaf area on
lite (1:1, vol/vol) in the greenhouse. Plants were used for disease each plant, control efficacy, and ED90 values were calculated as
control assays at 3 to 5 weeks after seeding. described above. For each pathosystem, the mean percentage of
Cucumber leaves infected by pathotype 3 of Pseudoperono- infected leaf area (pustule number for sunflower rust) and standard
spora cubensis (6) were collected from field-grown cucumbers at deviation of the mean were calculated. Analysis of variance was
Bar-Ilan Farm in spring 2001. Leaves were kept on wet filter performed followed by Fisher’s protected LSD test to establish
papers in 20-by-20-by-3-cm plastic trays at 20°C for 2 days (12 h significant differences (P = 0.05) in disease severities among
of light/day) to stimulate sporulation of the pathogen. Isolate 367 plants treated with different concentrations of each paste and be-
(A2 mating type, resistant to metalaxyl) (3) of Phytophthora infes- tween treated and untreated control plants for each pathosystem.
tans was kept and transferred on detached leaves of tomato. Thin-layer chromatography separation of paste. Paste
Sporangia of Pseudoperonospora cubensis and Phytophthora in- (500 mg, Keramim, June 2002) was dissolved in 0.5 ml of ace-
festans were collected with a fine brush into cold distilled water, tone. The solution was streaked on three thin-layer chromatography
the concentration was adjusted to 1 × 103 sporangia/ml, and the (TLC) plates (Silica 60, Merck 5725) and run in chloro-
suspensions were used for inoculation. form:methanol, 9:1 (vol/vol), to a distance of 17 cm. Based on
Sunflower leaves carrying urediniospores of Puccinia helianthi iodine vapor detection, six major colored regions (regions 1 to 6)
were obtained from Hazera Genetics, Brorim, Israel. Uredinio- were identified, corresponding to the following Rf values: 1 = 0
spores were suspended in water containing 0.01% Tween 20, their to 0.15, 2 = 0.15 to 0.43, 3 = 0.43 to 0.58, 4 = 0.58 to 0.78, 5 =
concentration was adjusted to 1 × 104 spores/ml, and the suspen- 0.78 to 0.84, and 6 = 0.84 to 094. Silica of each region was
sion was used for inoculation of sunflower plants. scraped and eluted in acetone. The acetone was evaporated and
B. graminis f. sp. tritici was obtained from the Institute of the weight of each component was determined. Components were
Cereal Research, Tel-Aviv University, and maintained on wheat emulsified in water (see above), diluted into 250, 125, and
seedlings (‘Sharon’). Fresh conidia produced on wheat seedlings 62.5 µg/ml, and applied to 10 tomato leaf disks (12 mm in diam-
were used for inoculation by gentle dusting over the test plants. eter) in a petri dish, five 10-µl droplets per disk. Droplets were
Activity of extracts dissolved in acetone. Pastes were dis- allowed to dry for 1 h and each leaf disk was inoculated with a
solved in acetone and diluted into four concentrations (2, 1, 0.5, 10-µl droplet of sporangial suspension (2,000 sporangia/ml) of
and 0.25%, wt/vol) as described above. Three replicate plants of Phytophthora infestans. Plates were incubated at 18°C in the dark
cucumber, potato, tomato, and sunflower and three pots having 10 for 16 h and then transferred to a 20°C growth cabinet as above.
wheat plants/pot were used per treatment. The solutions were Percentage of leaf disk area occupied with mycelia and sporangia
sprayed onto the upper leaf surface (both surfaces in wheat) of the of the pathogen was recorded 7 days after inoculation. Untreated
plants. Plants treated with acetone and untreated plants served as inoculated leaf disks and emulsifier-treated inoculated leaf disks
controls. At about 30 min after treatment, when the acetone had served as controls. Percentage of inhibition of late blight develop-
evaporated, treated plants and control plants were inoculated with ment was calculated relative to the untreated control inoculated
their respective pathogens. Inoculated plants, except wheat, were disks.
kept in a dew chamber for 12 to 16 h and then incubated in
growth chambers at 20°C (12 h of light/day, 100 µE m–2 s–1) for 6 RESULTS
to 10 days, depending on the pathosystem. Wheat plants were
placed in growth chambers immediately after inoculation. The Dry matter and paste contents. Dry matter content in fresh
percentage of infected leaf area in each plant was estimated leaves collected at Keramim in 2002 was 16, 22, and 25% in
visually. In sunflower, the number of pustules on each plant was April, June, and September, respectively. Dry matter content in
counted. Control efficacy was calculated as 100 (1 – X/Y), where fresh leaves collected at Bar-Ilan during 2003 ranged from 16.7 to
X = percent infected leaf area in treated plants and Y = percent 27.8%, depending on the time of harvest (Fig. 1A). It was lower
infected leaf area in acetone-control plants. The same calculation in leaves collected in April than in leaves collected from May to
was applied for sunflower, except pustule numbers per plant were October. In October, when plants started to bloom, dry matter
used for the calculation. Linear regression analysis was conducted content was higher than in July but similar to the other harvests,
between the log concentration of the extract (%) and probit of except April.
control efficacy (%) using SPSS 11.5 for Windows. Effective dose After extracting the dry leaves, the solvent mixture was evapo-
(concentration) required for 90% inhibition of disease (ED90) for rated leaving an oily, water-insoluble, green paste. Weighing the
each extract was calculated based on the linear regression equa- paste derived from 100 g of dry leaves enabled us to obtain the
tions. For each pathosystem, the mean percentage of infected leaf percentage of extractable paste in the dry leaves (paste yield). At
area (pustule number for sunflower rust) and the standard devi- Keramim, paste yield in April, June, and September was 9.0,
ation of the means of the seven monthly harvests were calculated 13.0, and 15.1, respectively. Paste yield obtained from dry leaves
with Super Anova. The mean ED90 values, the standard deviation collected at Bar-Ilan during 2003 ranged from 6.8 to 15.4% (Fig.
of the mean of the two harvests made each month for each 1B). The paste yield in leaves collected in April was lower than in
pathosystem, and those for all harvests for each pathosystem were leaves collected in the following 6 months.

Vol. 94, No. 10, 2004 1043

 Efficacy of pastes dissolved in acetone. Paste extracts derived late blight in potato, and powdery mildew in wheat ranged be-
from leaves collected at Keramim in April, June, and September tween 70 to 80% for paste extracts derived in April, compared
2002 were dissolved in acetone (1%, wt/vol) and tested for dis- with 90 to 95% for extracts derived in June or September (data
ease control. Mean control efficacy against late blight in tomato, not shown).
Figure 2 presents data for one extract derived from leaves col-
lected at Bar-Ilan on 15 June 2003. It reveals that the severity of
downy mildew of cucumber, late blight of potato or tomato, pow-
dery mildew of wheat, and rust of sunflower gradually decreased
when treated with increasing doses (0.25 to 2%, wt/vol) of paste
in acetone solution. At 0.25%, there were significant reductions in
disease severity relative to acetone-treated plants, and at a con-
centration of 1% the efficacy reached 90% or more. No differ-
ences in disease severity were observed between untreated inocu-
lated plants and acetone-treated inoculated plants (data not shown).
Similar experiments were conducted with all other pastes de-
rived from the field at Bar-Ilan during 2003. Disease severity data
were collected, percent efficacy was calculated, and ED90 values
were derived after log-probit transformation. Goodness of fit of
the linear regressions used to calculate the ED90 values ranged,
for the different harvests, between 0.889 and 0.997 for cucumber
downy mildew, 0.906 and 0.998 for potato late blight, 0.916 and
0.993 for tomato late blight, 0.871 and 0.999 for wheat powdery
mildew, and 0.838 and 0.997 for sunflower rust. The results (Fig.
3A to E) indicate differences between pastes and among patho-
systems. In most pathosystems, the efficacy of the paste obtained
in April was lower compared with the pastes obtained later in the
season. Mean ED90 values calculated from the data of Figure 3
are given in Figure 4. Mean ED90 values ranged from 0.68 to
1.02%, greater then the value of <0.18% for grape downy mildew
in a previous study (21). The order of sensitivity to the pastes, in
terms of mean ED90, was wheat powdery mildew < sunflower
rust < potato and tomato late blight < cucumber downy mildew
(Fig. 4).
Fig. 1. A, Dry matter content in fresh leaves of Inula viscosa harvested at bi-
Efficacy of paste emulsified in water. A paste, derived from
weekly intervals from April to October 2003. Values are means and standard leaves harvested in June 2002, was formulated with the aid of an
deviations of two harvests per month. B, Yield of paste extract in dry leaves of I. emulsifier and sprayed, at various concentrations, onto the foliage
viscosa harvested at biweekly intervals from the field during 2003 and dried of the test plants. The percentage of infected leaf area (or, for
at 60°C. Values are means and standard deviations of two harvests per month. sunflower rust, pustule numbers) for the various pathosystems is

Fig. 2. Percentage of leaf area infected by Pseudoperonospora cubensis, Phytophthora infestans, and Blumeria graminis f. sp. tritici in cucumber, potato and
tomato, and wheat, respectively, and pustules of Puccinia helianthi on each sunflower plant after treatment with either acetone alone or acetone solution
containing 0.25, 0.5, 1, or 2% paste of Inula viscosa leaves. DM/C, LB/P, LB/T, PM/W, and R/S stand for downy mildew/cucumber, late blight/potato, late
blight/tomato, powdery mildew/wheat, and rust/sunflower, respectively. The test extract of I. viscosa was prepared from leaves collected on 15 June 2003. Values
are means and standard deviations of three replicate plants (three pots of plants for wheat). For each pathosystem, means with the same letter are not significantly
different at P = 0.05 according to Fisher’s protected least significant difference test.

1044 PHYTOPATHOLOGY
presented in Figure 5. The data indicate ≈50% disease control of crop plants. They were effective not only against grape downy
with 0.125% of the emulsified paste. With 1% of the product, mildew caused by Plasmopara viticola as shown previously (21),
≈90% control was measured. The ED90 values for the different but also against cucumber downy mildew caused by Pseudo-
pathosystems were: downy mildew/cucumber, 1.01; late blight/ peronospora cubensis, late blight in potato and tomato caused by
potato, 0.97; late blight/tomato, 0.95; powdery mildew/wheat,
0.82; and rust/sunflower, 0.62%. The differences among the
various pathosystems were similar to those obtained with the ace-
tone solutions (Fig. 4). The percentage of infected leaf area
(pustule number for sunflower) in emulsifier-treated control
plants was similar to that in untreated control plants. The order of
sensitivity of the pathosystems to the emulsified paste extract was
similar to that of the acetone solution.
Activity of paste components. Data on the control efficacy of
late blight in tomato leaf disks of the components derived from
the paste (after TLC separation) are shown in Figure 6. Some
components were more active than others, while the emulsifier
itself enhanced disease development. Thus, at a concentration of
62.5 µg/ml, components derived from regions 1, 2, 3, 4, 5, and 6
of the TLC plates exhibited 0, 13, 41, 72, 82, and 78% inhibition
of the disease, respectively. At 250 µg/ml, only components 3, 4, Fig. 4. Effective concentration (percent, wt/vol) of Inula viscosa paste required
and 6 exhibited >85% inhibition, indicating that activity resides in for 90% inhibition (ED90) of Pseudoperonospora cubensis in cucumber
nonpolar compounds. It should be noted that components derived (DM/C), Phytophthora infestans in potato (LB/P), P. infestans in tomato
from a region may contain more than one compound. (LB/T), Blumeria graminis f. sp. tritici in wheat (PM/W), and Puccinia
helianthi in sunflower (R/S). Potted plants were treated with acetone alone or
acetone solution containing 0.25 to 2% pastes of I. viscosa leaves harvested in
DISCUSSION the field at biweekly intervals during April to October 2003. Values are means
and standard deviations of ED90 values for all harvests during the growth
The present study shows that extracts made from leaves of season. Means with the same letter are not significantly different at P = 0.05
I. viscosa possess broad-spectrum activity against foliar diseases according to Fisher’s protected least significant difference test.

Fig. 3. Effective concentration (percent, wt/vol) of Inula viscosa paste required for 90% inhibition (ED90) of A, Pseudoperonospora cubensis in cucumber, B,
Phytophthora infestans in potato, C, P. infestans in tomato, D, Blumeria graminis f. sp. tritici in wheat, E, and Puccinia helianthi in sunflower. Plants were
treated with acetone alone or acetone solution containing 0.25 to 2% pastes of I. viscosa leaves harvested in the field at biweekly intervals from April to October
2003. Values are means and standard deviations of ED90 values for two harvests per month.

Vol. 94, No. 10, 2004 1045

Phytophthora infestans, wheat powdery mildew caused by Yegen et al. (23) reported that aqueous extracts and essential
B. graminis, and sunflower rust caused by Puccinia helianthi. The oils of I. viscosa were antifungal in vitro. Muller-Riebau et al.
results corroborate earlier observations (4) showing that extracts (15), on the other hand, found only small amounts of antifungal
of I. viscose made with organic solvents are useful in disease essential oils or phenolics, concluding that the plant has no eco-
control. These findings may be significant to the agricultural in- nomical value for producing antifungal preparations. These authors
dustry when fungal strains resistant to site-specific fungicides also reported (14) that the main compounds in the essential oil are
prevail (5,8), as well as to organic farming where synthetic pesti- p-cymene and carvarol. Perez-Alonso et al. (17) found that the
cides are prohibited. yield of essentials oils of aerial parts was 0.2%. The major
The data showed that paste extracts dissolved in acetone are constituents were borneol, bornyl acetate, and isobornyl acetate.
effective in controlling diseases. Such paste extracts kept their Wollenweber et al. (22) identified 22 flavonoids in acetone ex-
high antifungal activity after storage of 3 years at room tem- tracts of I. viscosa leaves. Sanz et al. (19) analyzed methanol
perature (B. H. Ben-Daniel, unpublished data). However, because extracts of the foliage and identified flavonoids, sesquiterpene
acetone cannot be used in the field, we developed a technique to lactones, sesquiterpene acids, esters of 9-hydroxynerolidol, and
emulsify the paste obtained after extraction. The developed eudesmane acids. Grande et al. (9) isolated 10 triterpenoids, free
formulated product contains 34.4% paste. The emulsified product or esterified, from boiling acetone extracts of the aerial parts.
was effective in controlling all five diseases in potted plants,
whereas the emulsifier itself was ineffective at corresponding
doses. This EC product also was effective against downy mildew
on detached grapevine leaves and in grapevines grown in the field
(21). The 34.4% EC was effective after 1.5 years of storage at
room temperature and resisted heating or freezing for 2 weeks
(B. H. Ben-Daniel, unpublished data). Dried leaves of I. viscosa
stored at room temperature for 1 year produced an extract as
effective as newly harvested dry leaves or as dry leaves kept on
the bench for 4 years (unpublished data). These findings show
that the active ingredients are stable, which is valuable informa-
tion for industrial production of the extracts.
I. viscosa is well known for its strong odor and the stickiness of
its foliar parts. Extensive studies, therefore, have been conducted
by researchers to elucidate the nature or biological activity of its
water extracts, essential oils, and whole extracts in organic sol-
vents. Extractions were done using different techniques: in water
Fig. 6. Control of late blight on tomato leaf disks by components derived
(12,18), in boiling water (22), by autoclaving in water and parti-
from Inula viscosa. Dry paste was dissolved in acetone, applied to thin-layer
tioning with organic solvents (11,13), by distillation in water to chromatography plates, and separated into six regions. Silica scraped from
recover the essential oils (14), and by various organic solvents. each region was extracted with acetone and applied at various doses to the
These studies disclosed the presence of phenolics, flavonoids, leaf disks. Percent inhibition of disease development was calculated relative
terpenoids, sesquiterpene acids, sesquiterpene lactones, and other to untreated, inoculated disks. Standard deviations (data not shown) did not
compounds (1,2,4,7,9,15,19,20,22,23). exceed 12% of the corresponding mean value.

Fig. 5. Percentage of leaf area infected by Pseudoperonospora cubensis, Phytophthora infestans, and Blumeria graminis f. sp. tritici in cucumber, potato and
tomato, and wheat, respectively, and pustule numbers of Puccinia helianthi in each sunflower plant. Plants were treated with either water (untreated) or emulsion
in water containing 0.125, 0.25, 0.5, or 1% paste of Inula viscosa leaves or water emulsion containing the same rate of emulsifier only (blank control). DM/C,
LB/P, LB/T, PM/W, and R/S stand for downy mildew/cucumber, late blight/potato, late blight/tomato, powdery mildew/wheat, and rust/sunflower, respectively.
Values are means and standard deviations of three replicate plants (three pots of plants for wheat). For each pathosystem, means with the same letter are not
significantly different at P = 0.05 according to Fisher’s protected least significant difference test.

1046 PHYTOPATHOLOGY
 Chemical analyses conducted on our paste samples showed the LITERATURE CITED
presence of tomentosin, inuviscolide (sesquiterpene lactones)
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minimal inhibitory concentration values of 1.25 and 0.625 to synthesis in dermatophytes and Candida albicans. J. Ethnopharmacol.
2.5% (dry weight of leaves in water, wt/vol), respectively. Similar 71:479-482.
14. Muller-Riebau, F. J., Berger, B., and Yegen, O. 1995. Chemical-compo-
results were obtained by Ali-Shtayeh and Abu-Ghadeib (1). sition and fungitoxic properties to phytopathogenic fungi of essential oils
Our observations at Keramim in 2002 indicated that leaves har- of selected aromatic plants growing wild in Turkey. J. Agric. Food Chem.
vested in April contain less dry matter or paste and exhibited lower 43:2262-2266.
control efficacy compared with leaves harvested in June or Sep- 15. Muller-Riebau, F. J., Berger, B. M., Yegen, O., and Cakir, C. 1997. Sea-
tember. At Bar-Ilan we measured the (i) dry matter content in fresh sonal variations in the chemical compositions of essential oils of selected
leaves, (ii) paste (dry extract) yield, and (iii) efficacy of the extracts aromatic plants growing wild in Turkey. J. Agric. Food Chem. 45:4821-
4825.
in disease control. We assumed that fluctuations in these variables, 16. Oka, Y., Ben-Daniel, B. H., and Cohen, Y. 2001. Nematicidal activity of
if they occur, should be considered to maximize the economical powder and extracts of Inula viscosa. Nematology 3:735-742.
value of I. viscosa as a source for antifungal products. Therefore, 17. Perez-Alonso, M. J., Velesco-Negueruela, A., Duru, M. E., Harmandar,
extracts were prepared from leaves harvested at biweekly inter- M., and Vallejo, M. C. G. 1996. Composition of the volatile oil from the
vals from 1 April to 15 October 2003, the period of active growth aerial parts of Inula viscosa (L.) Aliton. Flavour Fragr. J. 11:349-351.
of I. viscosa in Israel. The results corroborate our preliminary ob- 18. Qasem, J. R., Al-Abed, A. S., and Abu-Blan, M. A. 1995. Antifungal
activity of clammy inula (Inula viscosa) on Helminthrosporium sati-
servations. They proved minor differences in dry matter content, vum and Fusarium oxysporum f. sp. lycopersici. Phytopathol. Mediterr.
paste yield, and efficacy of the different extracts, except for leaves 34:7-14.
collected in April, in which all the above values were lower 19. Sanz, J. F., Ferrando, C., and Marco, J. A. 1991. Oxygenated nerolidol
compared with the other harvests. For practical use, therefore, esters and eudesmane acids from Inula viscosa. Phytochemistry 30:3653-
harvesting can be done during almost the entire growth period. 3655.
Other data (data not shown) indicate that, after each harvest, the 20. Shtacher, G., and Kashman, Y. 1970. 12-carboxyeudesma-3,11(13)-diene.
A novel sesquiterpenic acid with a narrow antifungal spectrum. J. Med.
newly emerging shoots developing within ≈1 month also were suit- Chem. 13:1221-1223.
able for getting effective extracts. This means that harvesting can 21. Wang, W. Q., Ben-Daniel, B. H., and Cohen, Y. 2004. Extracts of Inula
be done repeatedly until late October, when plants start to flower. viscosa control downy mildew caused by Plasmopara viticola in grape-
The availability of herbal extracts, such as from I. viscosa, in vines. (Abstr.) Phytoparasitica 32:208.
the market may fulfill the need for a suitable product for organic 22. Wollenweber, E., Mayer, K., and Roitman, J. N. 1991. Exudate flavonoids
farming to combat destructive diseases in crop plants. of Inula viscosa. Phytochemistry 30:2445-2446.
23. Yegen, O., Berger, B., and Heitefuss, R. 1992. Investigations on the
fungitoxicity of extracts of 6 selected plants from Turkey against phyto-
ACKNOWLEDGMENTS pathogenic fungi. Z. Pflanzenkrank. Pflanzenschutz (J. Plant Dis. Prot.)
99:349-359. (In German)
This research was supported partly by a Fred and Barbara Kort Sino- 24. Ziv, O. 1996. Using extracts of Inula viscosa for controlling plant dis-
Israel Postdoctoral Fellowship to Dr. Wenqiao Wang. eases on fresh and dry post-harvest products. Phytoparasitica 24:154-155.

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