J. Agric. Sci. Mansoura Univ., 33 (4): 2659 - 2680, 2008
CONTROL OF GARDENIA LEAF SPOT AND BUD ROT
DISEASES USING SOME NATURAL PLANT OILS.
Mostafa, M.A. *, Effat
Nour El-houda A. Reyad*
A.
Zaher*,
A.H.
El-Shaer**
and
* Plant Pathol. Dept., Fac. of Agric., Cairo Univ.
** Plant Pathol. Res. Inst., A.R.C., Giza
ABSTRACT
This study was conducted to through light on the most important fungi affected
gardenia (Gardenia jasmenoides Ellis) plant with leaf spot and bud rots diseases and
the effect of some plant essential oils as safe management against these fungi in vitro
and in vivo. Isolation trials from infected gardenia plant tacked from Giza governorate
during 2005-2006 growing season revealed eleven fungal species related to eleven
genera. The isolates were differed in there frequency depending on the infected plant
part and the isolation periods, Botrytis cinerea, Alternaria alternata, Pestalotia
langloissii and Cladosporium sp. Were the most dominant fungi. These four isolates
were differed in there pathogenic capabilities depending on the infected plant part, B.
cinerea was exhibited the highest percentage of rotted buds while A. alternata and P.
langloissii were only infected the leaves. A. alternata was exhibited the highest disease
severity. Among twenty plant essential oils tested in vitro, Cumin (Cuminum cyminum)
oil was the most effective one, completely inhibited the mycelial growth of the tested
fungi at 500 ppm. concentration or more, while Anise (Pimpinella anisum), Peppermint
(Mentha piperita), Thyme (Thymus vulgaris), Clove (Syzigium aromaticum) and French
basil (Ocimum basilicum) oils at concentrations ranging between 750 and 1000 ppm
depending on the fungal species occupied the second position in this respect. In vivo
studies with cumin oil individually or in mixture with anise or clove oils were showed the
best treatment under artificial inoculation than under natural infection in reducing the
disease incidence. Generally spraying gardenia plant by cumin oil at 2500 ppm. mixed
with clove oil at 5000 ppm. concentration was the best treatment that significantly
decreased the disease incidence under greenhouse conditions.
INTRODUCTION
Production of ornamental plants is a considerable sector of the
economical agricultural income. Nowadays, the indoor plants became
necessary to overcome serious problems of air and environmental pollution,
particularly in the closed place or small apartments.
The Gardenia genus is considered one of the most important cutting
flower plants which includes over 200 species, with the most important ones
Gardenia jasminoides Ellis (native to china) and Gardenia thubergia L.F (White
gardenia, native to South Affrica). This genus is belonging to Rubiaceae
(Coffee) family.
The richly scented Gardenia jasminoides Ellis is suffering from several
diseases such as leaf spot (Barrett and Hardman, 1947; Shoemaker and
Straby; 1965; Gupta and Prased, 1983; kamal et al., 1983; Ciccaron, 1985;
Cappelli, 1996; Dreistadt, 2001; Kobayashi et al., 2003; Zheng-shiweii and
Lao-Chong, 2004 and Hilal, 2004), Flower blights and bud rots (Dimock, 1940;
Mullen and Jacobi, 2001; Pal et al., 1983; Dreistadt, 2001 and Hilal, 2004) and
stem canker (Calvino, 1939; Ghillini, 1940; Mc-Kenzie et al., 1940; Barret and
Mosafa, M. A. et al.
Hardman, 1947, Verneau, 1949 Buddin and Wakefield, 1938 and Hansen and
Barrett, 1938).
Gardenia leaf spot caused by Alternaria alternata and Pestalotia
langloissii as well as bud rots caused by Botrytis cinerea become to be serious
on G. jasminoides under Egyptian greenhouses, Particularly in moist
conditions where the disease can lead to heavy defoliation despite the
excessive and indiscriminate use of synthetic fungicides.
The excessive and indiscriminate use of synthetic fungicides are cause
many hazards to humans and animals due to their possible carcinogenicity,
teratogenicity, high and acute toxicity, long degradation periods and
environmental pollution (Lingk, 1991), Also, spraying these materials on the
foliages of gardenia plant results in a decrease in plant quality because of its
deposits on the leaves. So, the exploitation of natural substances such as
plant essential oils is urgently needed as alternatives to these synthetic
fungicides (Daferera et al., 2003), as they are easily decomposable, not
environmental pollutants and posses no residual or phytotoxic properties
(Tewari, 1990; Badei et al., 1996; Bishop and Thornton, 1997 and Tripathi et
al., 2002).
Essential oils are volatile, natural and complex compounds
characterized by a strong odour and are formed by aromatic plants as
secondary metabolites. Production of these oils by plants is believed to be
predominant a defense mechanism against pathogens (Oxenham, 2003) and
indeed, they have been shown to possess antifungal properties both in vitro
and in vivo (Wilson et al., 1997 and Bhaskara Reddy et al., 1998). The
complexity plant essential oils relates to their highly contents from natural
components that ranging between 20 and 60 components at quite different
concentrations. Among these large numbers of components there are two to
three components called the major components are find at fairly high
concentrations 20-70 % compared to others components present in trace
amounts (Bakkali et al., 2008). Although the major components are reflect
quite well the biophysical and biochemical features of the essential oils (Ipek et
al., 2005) it is difficult to correlate the fungitoxic activity to single compound or
class of compounds (Begamboula et al., 2004), where the synergistic or
antagonistic effect of one compound in minor percentage in the mixture must
be considered, as each of the essential oil components has its own
contribution on biological activity of the oil (Daferera et al., 2003). Generally,
inhibition of fungal growth by essential oils often involves induction of changes
in cell wall composition (Ghfir et al., 1997), plasma membrane disruption,
mitochondrial structure disorganization (de Billerbeck et al., 2001), and
interference with enzymatic reactions of the mitochondrial membrane, such as
respiratory electron transport, proton transport and coupled phosphoration
steps (Knobloch et al., 1989).
Plants have evolved physiological and biochemical mechanisms,
including increases in the activities of oxidative and reductive enzymes
associated with a biotic and biotic factors (Melo et al., 2006). This response
has been observed by several investigators related to plant defense (Farmer,
2001) and constitutes an evolutionary strategy of plants for defending
themselves against pathogens.
2660
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
The objective of this study is to through light on the most important fungi
affecting gardenia (Gardenia jasminoides Ellis) plants causing leaf spot and
bud rot diseases and the effect of some plant essential oils as safe
management against these fungi in vitro and in vivo.
MATERIALS AND METHODS
1-Isolation, purification and identification of the associated fungi.
Gardenia plants (Gardenia jasminoides Ellis) grown under greenhouse
conditions in commercial nurseries located at Giza governorate were suffered
from leaf spot and bud rots diseases all over the season. During February,
April, July and November of 2005-2006 growing season, diseased plant were
selected and transferred in their pots to the Lab of Plant Pathology Dept. Fac.
Agric. Cairo University.
Isolation trials were carried out during the previously mentioned
periods on potato dextrose agar medium (PDA). Spots with different sizes and
colours were carefully cut using sterilized forceps. The fragments were surface
sterilized using 1% sodium hypochlorite. Under aseptically conditions,
fragments were transferred to place onto the surfaces of sterilized PDA
medium in Petri-dishes (9 cm. in diam.). Petri-dishes were then incubated at
23° ±1°C for 3 days. The emerged fungi were picked up and subcultured onto
fresh PDA medium. Fungi were purified using hyphal tip or single spore
technique adopted by Dhingra and Sinclair (1985). Purified fungi were
identified according to their morphological characters using the keys given by
Ellis, (1971); Raper and Fennel, (1977) and O’Donnell, (1979).
Occurrence and frequency of fungi isolated at the end of the
incubation period were determined according to the following formula:
%X = N/T x 100
Where:
% X = frequency of the fungus.
N = Number of colonies for the fungus.
T = Total number of fungal colonies for the isolated fungi.
2- Pathogenicity testes:
Pathogenicity testes were conducted for the most dominant fungi
isolates namely, Botrytis cinerea, Alternaria alternata, Pestalotia langloissii and
Cladosporium sp. Each isolate was separately grown on PDA medium at 23°
±1°C for 7 days. A spore suspension was prepared by adding 10 ml. of distilled
water to each plate and tapering the spores using a camel hair brush. The
spore concentration was adjusted to 5x103 conidia/ml. using a haemicitomiter.
Under greenhouse conditions, where the degree of temperature was 25º±2 º C
and relative humidity (RH) was 65%, healthy gardenia plants (6-month old)
grown in pots 20 cm in diameter containing autoclaved peatmos were used in
this investigation. Before artificial inoculation, a drop of Tween 20 was added
to the spore suspension as a wetting agent. Spore suspension of any of the
four tested fungi was sprayed on both leaf surfaces and buds of the plant with
an atomizer and each plant received 20 ml of spore suspension. Treated
plants were covered with polyethylene bags to maintain high relative humidity
for 24h then removed. Control plants, were similarly treated only by sterile
2661
Mosafa, M. A. et al.
distilled water mixed with a drop of Tween 20. Three plants were used for
each particular treatment. Plants were observed daily for three weeks following
inoculation looking for leaf spot and bud rot symptoms.
Rotted buds on each treated plant were determined after 7 days
following inoculation period as percentage of rotted buds to the healthy one.
Whereas, for leaf spot determination, disease index was measured within
three weeks following inoculation. Areas of visible symptoms were scored for
disease index on a scale of 4 points as follows:
0 = no symptoms.
1 = few scattered lesions covering about 1-10% of the leaf.
2 = spots covering about 11-25% of the leaf.
3 = spots coalescing and covering about 26-50% of the leaf.
Disease index was converted according to the equation suggested by Baudion
(1988) as follows:
Disease index % = Σ n/N x 100
Where:
(n) Is the number of leaves in each numerical grade
(r) and (N) is the total number of inoculated leaves multiplied by the maximum
numerical grade (4).
3- Plant oils treatments:
A- Source of plant oils.
Several essential oils were tested for their antagonistic effects against
the three tested fungi i.e., B. cinerea, A. alternata, and P. langloissii the causal
agents of leaf spot and bud rots of gardenia plant either in vitro and in vivo.
The tested oils were from clove ( Syzigium aromaticum L.), anise ( Pimpinella
anisum ), peppermint ( Mentha piperita ), cumin ( Cuminum cyminum ),
coriander ( Coriandrum sativum ), French basil ( Ocimum basilicum ), local
basil (Ocimum kilimandscharium), caraway (Carum carvi), thyme (Thymus
vulgaris), fennel (Foeniculum vulgare), Egyptian geranium (Pelargonium
graveolens), sage (Salvia officinalis), rosemary (Rosmarinus officinalis),
chamomile (Ormenis mixta), parsley (Petroselinum crispum), marjoram
(Origanum vulgare), dill (Anethum graveolens), celery (Apium graveolens),
eucalyptus (Eucalyptus citriodora) and tagets (Tagetes patula). These plant
oils were obtained from Medicinal and Aromatic Pl. Res. St. El-Quanter ElKhairiah, Qulubyiah governorate, Hort. Res. Inst., Agric. Res. Center (ARC),
Giza, Egypt.
B- In vitro experiments:
1- In vitro effect of the individual plant oils on the linear growth of the
tested fungi:
The minimum inhibitory concentration of the tested plant oils against
the three tested fungi was conducted using a poisoned plate technique or the
agar dilution method described by Gulluce et al. (2003). The oils were added in
a separated mean to sterile melted PDA medium containing drop of Tween 20
to produce the concentrations 500, 750, 1000, 1500 and 3000 ppm. The
resulting PDA solutions were immediately poured into sterilized Petri-dishes (9
cm. in diam.) at the rate of 20 ml/plate. Dishes were inoculated at the center
with 4 mm mycelial disc cut from the periphery of 7- day-old culture of any of
the three tested fungi. The inoculated plates were incubated at the optimum
2662
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
temperature for each fungus, i.e. 20°C for B. cineria and 25°C for A. alternata
and P. langloissii. Three replicate dishes were used for each treatment. Plant
oil free PDA medium with drop of Tween 20 was used as control. The diameter
of the developed colonies was measured when the mycelial growth of the
fungus covered the plates of check treatment. The inhibition in mycelial growth
rate was calculated according to formula suggested by Deans and Svoboda
(1990) as follows:
I=C-T/C x100
Where:
(I) is the inhibition percentage of mycelial growth, (C) is the mean
colony diameter (mm) of the control set and (T) is the mean colony diameter of
treatment sets.
2-Effect of mixing cumin, clove, thyme, peppermint and anise oils on the
growth of the tested fungi:
Five of the most effective plant oils were selected to study their toxicity
on the tested fungi when mixed together, i.e. cumin (Cuminum cyminum),
thyme (Thymus vulgaris), anise( Pimpinella anisum ), peppermint ( Mentha
piperita ), and clove ( Syzigium aromaticum L.) oils, where the mixture was
conducted between each two oils at the rate of 1:1 (500:500 ppm.), 1:2
(250:500 ppm.) and 2:1 (500:250 ppm) concentrations. The toxicity of each
mixture was tested by the method mentioned above and described by Gulluce
et al. (2003). Three replicates were used for each treatment. Plant oil free PDA
medium with a drop of Tween 20 was used as control. Also, mycelial parts of
fungi which could not grow during the assay were transferred into sterile
essential oil-free PDA media and then observed for a week to determine the
fungicidal or fungistatic effect for each mixture of plant oils. The days needed
for mycelium reactivation for each fungus were also calculated.
3- Effect of cumin oil individually or in mixture with anise or clove oil on
percentage of mycelial growth and germinated sclerotia of Botrytis
cinerea.
Sclerotia of Botrytis cinerea the causal organism of gardenia bud rot
disease were dipped in cumin oil at 500 ppm. as individual treatment and on
the mixure of cumin and anise or clove oils at the rate of 1:2 (i.e. 250:500
ppm.) concentrations for 15 minutes. Sclerotia were removed with the help of
sterilized fine forceps, placed on sterilized blotting paper to remove access of
water and each sclerotium placed at the center of PDA medium. For control,
sclerotia were only dipped in sterilized water then placed on PDA medium.
Petri-dishes were incubated at the optimum temperature (20°C) for the fungus.
Three replicates were used for each treatment. The averages of the linear
growth in mm were calculated when the mycelium reached its maximum
growth in the check treatment.
On the other hand, the sclerotia were centrifuged with the three oils at
the same concentrations mentioned above as described by Zewani et.al.,
(2004) at 1000 rpm for 15 min. and decanted to remove mycelial fragments.
Sclerotia were removed with the help of sterilized fine forceps, placed on
sterilized blotting paper to remove access of water and then placed on PDA
medium. For control, sclerotia were centrifuged at 1000 rpm for 15 min. with
sterilized distilled water only. Five sclerotia were kept for each treatment and
2663
Mosafa, M. A. et al.
replicated three times. Percentages of germinated sclerotia were calculated
24h following incubation at the optimum temperature i.e. 20°C.
C-In vivo experiments:
1-Effect of cumin oil individually or in mixture with anise or clove oils on
controlling bud rot caused by B. cinerea and incidence of leaf spot
disease caused by A. alternata and P. langloissii.
Healthy Gardenia plants (6-month old) grown in sterilized pots (25cm in
diam ) containing peatmos were used to study the effect of cumin oil
individually and mixurally with anise or clove oil on controlling the infection
caused by the three tested fungi under greenhouse conditions. The tested oils
were diluted to 5000 ppm for cumin oil treatment and 2500:5000 ppm for the
mixtures, where the concentration of cumin oil in the mixture was 2500 ppm,
meanwhile both clove and anise oils were mixed by 5000 ppm for each. The
tested oils sprayed onto the upper leaf surfaces to run-off using an atomizer
24h before inoculating the plants with a spore suspension of each fungus
(5.0x10³ conidia/ml).Control treatment consisted of sterilized distilled water
containing 0.5 % Tween 20. The percentage of rotted buds was assessed 8
days following the inoculation. Meanwhile, the disease index of leaf spot
symptoms was assessed 3 weeks following inoculation. Three pots were used
for each treatment.
2-Effect of cumin oil individually and in a mixture with anise or clove oil
on controlling bud rot and leaf spot diseases under natural infection
under greenhouse conditions.
To study the effect of cumin oil individually or in mixture with anise or
clove oil under greenhouse conditions, healthy Gardenia plants (6-month old)
grown in sterilized pots (25cm in diam.) containing peatmos were divided into
three groups. At the beginning of March ,2007 the first group sprayed by cumin
oil at 5000 ppm concentration, the second group sprayed by a mixture of
cumin oil at 2500 ppm and anise oil at 5000 ppm concentration and the third
group sprayed by a mixture of cumin oil at 2500 ppm and clove oil at 5000
ppm concentration. The plants were sprayed randomly by atomizer each week
through 2007 season. Three replicate pots were used for each treatment.
Control treatment consisted of healthy plants sprayed with sterilized distilled
water containing 0.5 % Tween 20. The rotted buds were observed at May
2007 and the percentage of rotted buds was calculated as mentioned under
pathogenicity test. The resulting leaf spots symptoms were observed at
September 2007 and the disease index was calculated as mentioned before.
RESULT AND DISCUSSION
1-Isolation, purification and identification of the associated fungi.
In the present study (Table 1) Isolation trails, from naturally infected
gardenia plant by leaf spot and bud rot symptoms collected from greenhouses
located at Giza governorate resulted in the presence of eleven fungal species
belonged to eleven fungal genera. These fungi were identified as Botrytis
cinerea, Alternaria alternata, Pestalotia langloissii, Cladosporium sp.,
Stemphylium sp., Phomopsis sp., Myrothecium roridum, Coniotherium sp.,
Fusarium oxysporum., Botryodiplodia sp. and Trichoderma viride. These
2664
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
isolates were differed in their occurrence and frequencies according to the
infected plant part and the seasonal isolation periods. Generally, B. cinerea, A.
alternata, P.langloissii and C. sp. were the most prevailing fungi. B. cinerea
was isolated only from the rotted buds formed in spring season at the rate of
77%, where it was recorded the highest frequency of occurrence during this
period. A. alternata, P.langloissii and C. sp. were isolated from both infected
leaves and buds through all the isolation periods except for A. alternata which
not isolated in spring season. The heights frequency of C. sp. was recorded in
winter season at the rate of 50% and 75% from infected leaves and buds,
respectively followed by spring season by 57.1% and 15.4% from leaves and
buds, respectively. Meanwhile, the highest frequency of A. alternata and P.
langloissii were recorded in summer season. The corresponding percentages
were 40% and 15.4% for A. alternata and 30% and 30.8% for P. langloissii
from infected leaves and buds, respectively.
Table 1: Occurrence and frequency of fungi isolated from gardenia plants
suffering from leaf spot and bud rot diseases through 20052006 growing season.
Isolated fungi
Botrytis cinerea
Alternaria
alternata
Pestalotia
langloissii
Cladosporium sp
Myrothecium
roridum
Coniotherium sp
Stemphelium sp
Fusarium
oxysporum
Botryodiplodia sp
Phomopsis sp
Trichoderma
viridae
Spring
Leaves Buds
00.0
77.0
% frequency / season
Summer
Autumn
Leaves Buds Leaves Buds
00.0
00.0
00.0
00.0
Winter
Leaves Buds
00.0
00.0
00.0
00.0
40.0
15.4
20.0
00.0
16.7
00.0
14.6
00.0
30.0
30.8
20.0
00.0
16.7
25.0
57.1
15.4
00.0
23.2
35.0
100.0
50.1
75.0
14.3
00.0
00.0
00.0
00.0
00.0
16.7
00.0
00.0
14.0
00.0
00.0
10.0
00.0
00.0
00.0
20.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
15.0
00.0
05.0
00.0
00.0
00.0
00.0
00.0
7.6
00.0
00.0
05.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
00.0
30.8
00.0
00.0
00.0
00.0
According to the available literature, the recorded causal organisms of
leaf spot and/or bud rot diseases of Gardenia jasminoides Ellis in Egypt were
Fusarium solani, Nigrospora sp., Rhizoctonia solani on the flowers and
Myrothecium roridum on the leaves (Hilal, 2004), but in other countries
Numerous investigators have shown that leaf spot and bud rots diseases of
Gardenia jasmenoides Ellis are caused by different fungi such as Botrytis
primer (Mullen and Jacobi, 2001); Botrytis cinerea (Dimock, 1940 and
Dreistadt, 2001); Alternaria alternata (Gupta and Prasad, 1983); Pestalotia
langloissii (Shoemaker and Straby, 1965); Pestalotia spp. (Gonsalves and
Ferreira., 2002); Myrothecium roridum (Barrett and Hardman, 1947; Cappelli,
1996 and Dreistadt, 2001); Rhizoctonia spp. (Dreistadt, 2001); Septoria
gardeniae (Kobayashi et al., 2003); Cercosporidium okinawaense (Kobayash
2665
Mosafa, M. A. et al.
et al., 2002); Phyllosticta gardiniicola (Zheng and Lao, 2004 and Kamal et al.,
1938); Phyllosticta sp. (Dreistadt, 2001); Mycosphaerella luzonensis
(Kobayashi et al., 1988 and Colletotrichum gloeosporides (Ciccaron, 1985; and
Zheng-Shiweii and Lao-Chong, 2004). This higher numbers of the recorded
fungi that associating with gardenia plant may be attributed to the
physiobiochemical processes in the plant.
2- Pathogenicity testes:
Pathogenicity tests by The most prevailing fungi isolated from
gardenia plants namely, B. cinerea, A. alternate, P. langloissii and
Cladosporium sp., indicated that these isolates were differed in there
pathogenic capabilities to leaves and buds of the plant (Table 2.) The
pathogenic fungi were Botrytis cinerea, Alternaria alternata and Pestalotia
langloissii but Cladosporium sp was non-pathogenic. B. cinerea, A. alternata
and P.langloissii were specialized in their pathogenic capabilities depending on
the infected plant part. B. cinerea was only pathogenic to the plant buds,
where it exhibited the highest percentage of rotted buds, being 41, 7% .
Meanwhile, A. alternata and P. langloissii were pathogenic to the leaves. A.
alternata exhibited the highest disease index, being 47.8% while, P. langloissii
was the lowest one in this respect, being 21.1%.
Table (2): Pathogenicity tests using Botrytis cinerea., Alternaria alternata,
Pestalotia langloissii and Cladosporium. sp.
Fungi tested
Botrytis cinerea
Alternaria alternata
Pestalotia langloissii
Cladosporium sp
% Disease index of Leaf
spots
00.0
47.8
21.1
00.0
% infection
by Bud rots
41.7
00.0
00.0
00.0
3- Plant oils treatments:
A- In vitro experiments:
1-In vitro effect of the individual plant oils on the linear growth of the
tested fungi:
Data obtained (Tables 3,4 and 5) clear that as the concentration of
each oil increase, linear growth of the tested fungi decreased, and this effect
differed according to the oil type. Cumin oil was the most effective one,
completely inhibited the mycelial growth of the tested fungi at 500 ppm
concentration or more, while clove, anise, thyme, coriander, French basil and
peppermint oils at concentrations ranged from 750 to 1500 ppm according to
the fungus type occupied the second position in this respect. Where, clove,
anise, thyme and peppermint oils at 750, 1000, 1500 and 1500 ppm
concentrations, respectively completely inhibited the mycelial growth of A.
alternata. anise, peppermint and thyme oils at 750 ppm concentration as well
as clove, French basil and geranium oils both at 1000 ppm also, rosemary oil
at 1500 ppm concentration completely inhibited the mycelial growth of P.
langloissii. Meanwhile, anise, thyme, peppermint, clove, coriander and French
basil oils at 1000 ppm concentration completely inhibited the mycelial growth
of B. cinerea.
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J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
Table 3: Linear growth (mm) and percentage of reduction in mycelial
growth of B. cinerea on PDA medium mixed with different
concentrations of 20 plant essential oils and incubated at 20° C
for 6 days.
Oils
(O)
tested
linear growth (mm) and (%) reduction in mycelium growth at (ppm)
concentration of essential oils
500 Red% 750 % Red 1000 % Red 1500 % Red 3000 % Red Mean
Cuminum
00.0 100.0
cyminum
Pimpinella
24.6 72.7
anisum
Mentha
13.0 85.6
piperita
Thymus
29.8 66.9
vulgaris
Syzigium
39.1 57.0
aromaticum
Coriander
52.0 42.2
sativum
Ocimum
68.0 24.4
basilicum
Carum carvi 45.8 49.1
O.kilimandsc
62.2 30.9
harium
Foeniculum
61.0 32.2
vulgare
Pelargonium
90.0 00.0
graveolens
Salvia
90.0 00.0
officinalis
Rosmarinus
90.0 00.0
officinalis
Origanum
90.0 00.0
vulgare
Petroselinu
90.0 00.0
m crispum
Anethum
90.0 00.0
graveolens
Ormenis
43.3 51.9
mixta
Apium
90.0 00.0
graveolens
Eucalyptus
90.0 00.0
citriodora
Tagets
90.0 00.0
patula
(Check )
Mean
62.6
LSD at 0.05 for:
Oils tested (O):
Concentration (C):
O×C:
00.0
100.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
18.0
80.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
100.0
02.6
82.1
00.0
100.0
00.0
100.0
00.0
100.0
09.2
19.8
78.0
00.0
100.0
00.0
100.0
00.0
100.0
11.8
37.7
58.1
00.0
100.0
00.0
100.0
00.0
100.0
18.0
43.2
52.0
00.0
100.0
00.0
100.0
00.0
100.0
22.2
27.3
69.7
19.0
79.1
14.3
84.1
00.0
100.0
21.3
56.7
37.0
23.5
74.0
15.0
73.3
00.0
100.0
31.5
51.8
42.4
33.0
63.3
21.0
76.7
00.0
100.0
33.4
45.0
50.0
29.3
64.4
17.0
81.1
00.0
100.0
36.3
90.0
00.0
37.0
58.9
18.7
79.2
00.0
100.0
47.1
90.0
00.0
74.0
17.8
61.0
32.2
00.0
100.0
27.0
90.0
00.0
43.7
51.4
33.0
63.3
00.0
100.0
51.3
90.0
00.0
68.3
24.1
41.7
58.3
18.2
79.8
61.6
90.0
00.0
40.3
55.2
26.8
70.2
13.0
85.0
52.0
37.5
58.3
29.0
67.8
23.7
73.7
20.7
77.0
90.0
00.0
72.0
20.0
64.8
28.0
46.5
48.5
72.7
90.0
00.0
90.0
00.0
71.0
21.1
25.2
72.0
73.2
90.0
00.0
90.0
00.0
53.3
21.1
40.8
28.3
68.6
16.1
90.0
35.2
54.5
23.9
12.0
30.8
----
0.3
0.2
0.8
On the other hand, caraway, local basil, fennel, marjoram, rosemary and
geranium oils at 3000 ppm completely inhibited the mycelial growth of the
tested fungi. Meanwhile, celery, parsley, chamomile, eucalyptus, tagets oils
2667
Mosafa, M. A. et al.
were the lowest activity. Where each of them only resulted in reduced of
mycelial growth of the tested fungi till the highest tested concentration 3000
ppm. These results are in harmony with several workers. Maruzzella (1962)
arranged the active parts of volatile oils according to the antimicrobial activities
in the decreasing orders as follows: Aldehyde, Phenols, Alcohols, Ketons and
Hydrocarbons. Jaspal and Tripathi (1999) mentioned that the pure essential
oils completely inhibited the mycelial growth of many pathogenic fungi, and the
fungal sensitivity to the previous essential oils differed in their effect from one
fungus to another, this might be due to the capability of essential oils to
penetrate into the fungal cells. According to Aligianise et al. (2001), the activity
of the plant essential oils was divided depending on the minimum inhibitory
concentration (MIC) to three divisions i.e. strong (MIC more than 500 ppm),
Moderate ( MIC from 600 ppm to 1600 ppm ) and weak (MIC more than 1600
ppm). So, the results obtained from the present study of cumin oil on the fungi
tested lead to group the oil into a strong effect category, where it is completely
inhibited the mycelial growth of the tested fungi at 500 ppm concentration The
inhibitory effect of cumin oil might be attributed to the presence of the
cuminaldehyde. Anise, peppermint, clove, coriander, thyme and French basil
oils were showed a moderate affect. Where, they completely inhibited the
mycelial growth of the fungi tested at concentrations ranging from 750 to 1500
ppm. ,except for coriander oil which showed a week effect only on A. alternata
where it is completely inhibited the mycelial growth of the fungus at 3000 ppm
concentration., This might be due to the presence of the great amount of
phenolic and alcohols substances like thymol in thyme oil, eugenol in clove oil,
menthol and carvacrol besides menthyl acetate ester in peppermint oil,
chavicol beside linalool alchohol, methyl chavicol and camphen compound in
French basil oil (Mohamed et. al., 2003; El-Baroty, 1988 and Moussa, 1998).
The other tested oils i.e. caraway, local basil, fennel, marjoram, rosemary,
geranium, celery, parsley, chamomile, eucalyptus, tagets oils were the lowest
activity. Where each of them only resulted in a reduction of the mycelial
growth of the tested fungi till the highest tested concentration 3000 ppm.,
except for rosemary and geranium oils which showed a moderate affect only
on P. langloissii where they completely inhibited the mycelial growth of the
fungus at 1500 and 1000 ppm concentration respectively. This week activity of
the previously mentioned oils might be due to their poorness in phenolic
compounds for example local basil contains the same compounds in French
basil but in small quantities also, marjoram oil is very poor in its phenolic
compounds except cineol compound which act as antifungal agent in a great
amounts reaching to 30.1% but it was poor in other phenolic compounds (ElBaroty, 1988; Moussa, 1998; Bhaskara Reddy et al., 1998 and Arras and Usai,
2001).The MIC and toxicity concentrations of the essential oils varied from
study to study and this is probably due to the different methods of extraction of
the essential oils and different sensitivity of the tested fungi (Saikaia et al.,
2001).
2668
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
Table
4:
Oils tested
(O)
Cuminum
cyminum
Pimpinella
anisum
Mentha
piperita
Thymus
vulgaris
Syzigium
aromaticum
Coriander
sativum
Ocimum
basilicum
Carum carvi
O.kilimandsc
harium
Foeniculum
vulgare
Pelargonium
graveolens
Salvia
officinalis
Rosmarinus
officinalis
Origanum
vulgare
Petroselinum
crispum
Anethum
graveolens
Ormenis
mixta
Apium
graveolens
Eucalyptus
citriodora
Tagets patula
(Check )
Linear growth (mm) and percentage of reduction in
mycelialgrowth of A. alternata on PDA medium mixed with
different concentrations of 20 plant essential oils and
incubated at 25° C for 10 days.
linear growth (mm) and (%) reduction in mycelium growth at (ppm)
concentration of essential oils
500 Red% 750 % Red 1000 % Red 1500 % Red 3000 % Red Mean
00.0 100.0 00.0 100.0 00.0 100.0 00.0 100.0 00.0 100.0 00.0
45.0
50.0
20.5
77.2
00.0
100.0
00.0
100.0
00.0
100.0
13.1
32.5
63.9
27.8
69.1
24.5
72.8
00.0
100.0
00.0
100.0
17.0
36.2
59.8
60.5
32.8
73.3
18.6
00.0
100.0
00.0
100.0
07.0
34.7
61.4
00.0
100.0
00.0
100.0
00.0
100.0
00.0
100.0
34.0
68.0
24.4
66.3
26.3
33.2
63.1
12.6
86.0
00.0
100.0
36.0
37.0
18.9
65.0
27.8
56.7
37.0
00.0
100.0
00.0
100.0
40.0
72.7
60.7
19.2
32.6
67.8
55.8
24.7
38.0
59.2
43.7
34.2
51.4
37.4
23.0
58.4
74.4
00.0
00.0
100.0
100.0
37.0
79.5
11.7
62.8
30.2
56.2
37.6
23.0
72.1
00.0
100.0
50.0
90.0
00.0
61.7
31.4
52.8
41.3
44.0
51.1
00.0
100.0
75.0
90.0
00.0
90.0
00.0
73.8
18.0
66.0
26.7
54.7
39.2
74.0
90.0
00.0
84.2
06.4
74.5
17.2
61.0
32.2
53.0
42.2
58.3
90.0
00.0
90.0
00.0
60.7
32.6
50.8
43.6
00.0
100.0
43.0
67.0
25.6
54.0
40.0
43.5
51.7
28.0
68.9
20.8
76.9
39.4
65.2
27.6
63.2
29.8
59.3
34.1
42.0
53.3
22.8
74.7
51.0
56.2
37.6
42.3
53.0
37.0
58.9
33.0
63.3
28.3
73.6
59.0
77.3
14.1
68.8
23.6
67.0
25.6
53.2
40.9
27.8
69.1
80.0
90.0
00.0
90.0
00.0
90.0
00.0
71.2
20.9
57.7
35.9
50.0
57.7
36.0
57.2
36.4
55.8
38.0
90.0
50.1
43.8
51.3
34.5
61.7
34.0
Mean
61.0
LSD at 0.05 for:
Oils tested (O):
Concentration (C):
O×C:
60.0
0.2
0.1
0.8
2669
34.1
19.0
40.0
-----
Mosafa, M. A. et al.
Table 5: Linear growth (mm) and percentage of reduction in mycelial
growth of P.langloissii on PDA medium mixed with different
concentrations of 20 plant essential oils and incubated at 20° C
for 6 days.
Oils tested
(O)
linear growth (mm) and (%) reduction in mycelium growth at (ppm)
concentration of essential oils
500 Red% 750 % Red 1000 % Red 1500 % Red 3000 % Red Mean
Cuminum
00.0 100.0 00.0 100.0 00.0 100.0
cyminum
Pimpinella
43.8 51.3 00.0 100.0 00.0 100.0
anisum
Mentha
45.7 49.2 00.0 100.0 00.0 100.0
piperita
Thymus
16.3 82.0 00.0 100.0 00.0 100.0
vulgaris
Syzigium
36.3 60.0 31.5 65.0
00.0 100.0
aromaticum
Coriander
19.8 78.0 11.3 87.4
00.0 100.0
sativum
Ocimum
65.5 27.2 55.3 38.6
00.0 100.0
basilicum
Carum
63.0 30.0 52.5 41.7
28.0
68.9
carvi
O.kilimand
63.0 30.0 48.7 45.9
39.6
56.0
scharium
Foeniculu
65.8 26.9 53.5 40.6
43.7
51.4
m vulgare
Pelargonium
56.3 37.4 51.2 43.1
00.0 100.0
graveolens
Salvia
90.0 00.0 69.7 22.3
53.5
40.6
officinalis
Rosmarinus
71.0 21.1 67.3 25.2
66.0
26.7
officinalis
Origanum
59.5 33.9 48.7 45.9
46.7
48.1
vulgare
Petroselinu
m crispum 21.3 76.3 19.2 79.0
17.0
81.1
Anethum
51.0 43.3 31.3 65.2
27.7
69.2
graveolens
Ormenis
29.3 67.4 25.0 72.2
23.3
74.1
mixta
Apium
65.2 27.6 49.0 46.0
55.2
61.0
graveolens
Eucalyptus
90.0 00.0 90.0 00.0
74.0
17.8
citriodora
Tagets
90.0 00.0 90.0 00.0
61.7
31.4
patula
(Check )
90.0
Mean
53.0
42.0
27.0
LSD at 0.05 for:
Oils tested (O):
0.2
Concentration (C):
0.1
O×C:
0.5
2670
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
09.0
00.0
100.0
00.0
100.0
09.1
00.0
100.0
00.0
100.0
03.3
00.0
100.0
00.0
100.0
13.6
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
24.2
16.4
81.8
00.0
100.0
29.0
23.0
74.4
00.0
100.0
22.3
27.0
70.0
00.0
100.0
33.0
00.0
100.0
00.0
100.0
22.0
17.0
81.1
00.0
100.0
31.0
00.0
100.0
00.0
100.0
17.0
36.4
59.6
00.0
100.0
26.0
14.7
83.7
11.0
87.8
24.0
16.2
82.0
07.8
91.3
38.4
21.3
76.3
19.0
78.9
72.0
23.5
73.9
19.2
78.7
70.0
63.8
29.1
42.0
53.3
41.0
56.0
37.8
46.8
48.0
70.3
14.4
11.2
----
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
2 -Effect of mixing cumin, clove, thyme, peppermint and anise oils on the
growth of the tested fungi:
The mixture between the tested oils completely inhibited the mycelium
growth of the tested fungi except for the mixture of (anise & thyme), (clove &
thyme), (clove & peppermint) and (thyme & peppermint) where they only
resulted in the reduction in mycelial growth of A. alternata from 90 mm in the
absence of oil (check) to 85.6; 77.8; 71.1 and 79.4 mm, respectively. (Table 6).
Boyraz and Ozcan (2006), mentioned that combinations of hydrosol, oleoresin,
ground material and essential oils may provide an efficacious mixture for the
inactivation of pathogenic and spoilage microorganisms in plant and foods.
Table (6): Effect of the tested plant oils mixtures on the linear growth of
the tested fungi.
B. cinerea
Linear
growth % Red.
(mm)
Oils tested (O)
Cuminum cyminum &
Pimpinella anisum
Cuminum cyminum &
Syzigium aromaticum
Cuminum cyminum &
Thymus vulgaris
Cuminum cyminum &
Mentha piperita
Pimpinella anisum &
Syzigium aromaticum
Pimpinella anisum &
Thymus vulgaris
Pimpinella anisum &
Mentha piperita
Syzigium aromaticum
Thymus vulgaris
Syzigium aromaticum &
Mentha piperita
Thymus vulgaris &
Mentha piperita
Cuminum cyminum &
Syzigium aromaticum
Cuminum cyminum &
Thymus vulgaris
Cuminum cyminum &
Pimpinella anisum
Mean
&
Fungi
A. alternata
P. langloissii
Linear
Linear
Mean
growth % Red. growth % Red.
(mm)
(mm)
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
13.0
85.6
00.0
100.0
04.3
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
20.0
77.8
00.0
100.0
06.7
00.0
100.0
26.0
71.1
00.0
100.0
08.7
00.0
100.0
18.5
79.4
00.0
100.0
06.2
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
100.0
00.0
100.0
00.0
100.0
00.0
00.0
----
07.8
----
00.0
----
----
3- Fungicidal and fungistatic effects of the tested oils individually and in
mixture on the tested fungi:
Cumin oil as individual treatment had a fungicidal effect on the tested
fungi. Meanwhile, the other individual oils tested had a fungistatic effect and
the days needed for mycelium reactivation ranged from 1 to 4 days according
to the fungus type. (Table 7). On the other hand the mixture of cumin oil at 500
ppm with any of the tested oils at 500 ppm gave a fungicidal effect on the
2671
Mosafa, M. A. et al.
tested fungi except for A. alternata where in case of the treatment with the
mixture of (cumin & thyme) and (cumin &peppermint) a fungistatic effect was
obtained and the mycelium reactivated through four days. this might be related
to the differences in the ultrasruction of the fungus conidia .Jaspal and Tripathi
(1999), mentioned that the pure essential oils completely inhibited the mycelial
growth of many pathogenic fungi and the fungal sensitivity to the previous
essential oils from one fungus to another, this also might be due to the
capability of essential oils to penetrate into the fungal cell.
Table 7. Fungicidal and fungistatic effects of the tested oils as individual
treatment at 500ppm on the growth of the fungi tested.
fungicidal and fungistatic effect of the oils / days on
mycelium reactivation
B. cinerea
A. alternata
P. langloissii
Cuminum cyminum
+
+
+
Pimpinella anisum
±
3*
±
2
±
2
Mentha piperita
±
1
±
3
±
3
Thymus vulgare
±
3
±
2
±
2
Syzigium aromaticum
±
3
±
3
±
3
Coriandrum sativum
±
4
±
2
±
1
Ocimum basilicum
±
4
±
2
±
2
Carum carvi
±
2
±
3
±
3
O. kilimandscharium
±
2
±
2
±
2
Foeniculum vulgare
±
4
±
3
±
2
Pelargonium graveolens
±
2
±
1
±
1
Salvia officinalis
±
1
―
1
±
1
Rosemarinus officinalis
±
2
―
1
±
1
Origanum vulgare
±
1
―
1
―
1
Petroselinium crispum
―
1
―
1
―
1
Anethum graveolens
―
1
―
1
―
1
Ormenis mixta
―
1
―
1
―
1
Apium graveolens
―
1
―
1
―
1
Eucalyptus citriodora
―
1
―
1
―
1
Tagetes patula
―
1
―
1
―
1
(+) Fungicidal effect
(±) Fungistatic effect
(―) No effect
* mycelium reactivation after 1-4 days
Plant oils treatments
On the other hand, reduced the concentration of cumin oil to 250 ppm
and fixed the concentration of any of the tested oils in the mixture at 500 ppm
had a fungicidal effect only in case of the mixture of (cumin & anise) and
(cumin & clove) oils on both B. cinerea and P. langloissii. Meanwhile, cumin oil
at 250 ppm mixed with clove oil at 500 ppm had a fungicidal effect on and A.
alternata. The other mixtures had a fungistatic effect on the tested fungi. While,
fixed the concentration of cumin oil at 500 ppm and reduced the concentration
of any of the tested oils in the mixture to 250 ppm had a fungicidal effect on the
tested fungi (Table 8). Anderson et al. (1994), mentioned that the fungicidal
activity of some essential oils constitutes such as trans-2-hexanal and citral
aldehydes may be ascribed to the high electrophilic properties of the carponyl
group adjacent to the double bond that make these compounds particularly
reactive with neucleophiles, such as protein sulfhydryl and amino groups of the
pathogen. Feng and Zheng (2007) mentioned that essential oils inhibited the
2672
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
growth of fungi either temporarily (fungistatic) or permanently (fungicidal).
Kishnora et al. (2007) reported that most essential oils are fungistatic effect
than fungicidal. In this study, the fungicidal effect was only recorded for the
cumin oil which contains cuminaldehyde as the major component. Also, the
mixtures containing this oil with any of anise or clove oil which both rich in their
content from phenolic compounds. So, this might be explained the fungicidal
effect of the tested oils on the fungi tested.
Table (8): Fungicidal and fungistatic effect of the most effective oils used
by 250:500ppm and 500:250ppm concentrations on the tested
fungi.
Plant oil treatment
C1
Cuminum
cyminum
C2
Cuminum cyminum
Pimpinella anisum
Mentha piperita
Thymus vulgaris
Syzigium aromaticum
------+
±
±
±
Cuminum cyminum
Pimpinella anisum
Mentha piperita
Thymus vulgaris
Syzigium aromaticum
------+
±
±
+
Cuminum cyminum
+
Pimpinella anisum
+
Mentha piperita
±
Thymus vulgaris
±
Syzigium aromaticum
+
C1=500ppm
C2=250ppm
(+) Fungicidal effect
Pimpinella
Mentha
Thymus
anisum
piperita
vulgaris
C2
C2
C2
Botrytis cinerea
+
+
±
------±
±
±
-----±
±
±
-----±
±
±
Alternaria alternata
+
+
±
------±
±
±
-----±
±
±
-----±
±
±
Pestalotia langloissii
+
±
±
------±
±
±
-------±
±
±
------±
±
±
(±) Fungistatic effect
Syzigium
aromaticum
C2
+
±
±
±
------+
±
±
±
-----+
±
±
±
---------
(―) No effect
4- Effect of cumin oil individually or in mixture with anise or clove oil on
percentage of
mycelial growth and germinated sclerotia of B.
cinerea.
Cumin oil at 250 ppm mixed with clove oil at 500 ppm concentration
recorded the heighst percent in reduced the mycelial growth raised from
sclerotia and its germination followed by cumin oil at 500 ppm concentration.
Meanwhile, cumin oil at 250 ppm mixed with anise oil at 500 ppm
concentration had the lowest one in this respect (Table 9and 10).
2673
Mosafa, M. A. et al.
Table (9): Effect of individual treating PDA medium with cumin oil alone
or in a mixture with anise or clove oil on the germination
percentage of sclerotia of B. cinerea, the causal pathogen of
gardenia bud rot disease.
Plant oil treatment
Cuminum cyminum
Cuminum cyminum
&
Pimpinella anisum
Cuminum cyminum
&
Syzigium aromaticum
(Check)
Mean
LSD at 0.05
Conc.
(p p m )
500
250
+
500
250
+
500
--------
Germinated sclerotia (%)
6 6 .7
5 3 .3
4 0 .0
1 0 0 .0
65.0
2.7
Table (10): Effect of cumin oil individually or in a mixture with anise or
clove oil on percentage of reduction in mycelial growth of
sclerotia of B. cinerea the causal pathogen of gardenia bud
rot disease.
Plant oil treatment
Cuminum cyminum
Cuminum cyminum
&
Pimpinella anisum
Cuminum cyminum
&
Syzigium aromaticum
(Check)
Mean
LSD at 0.05
Con. (ppm)
500
250
+
500
250
+
500
---------------
Linear growth (mm)
5 3 .4
% Red.
4 0 .7
6 8 .3
2 4 .1
3 4 .0
6 2 .2
90
31.8
1.6
00.0
---------
B-In vivo experiments:
1- Effect of cumin oil individually or in mixture with anise or clove oils on
controlling bud rot caused by B. cinerea and incidence of leaf spot
disease caused by A. alternata and P. langloissii.
The experiment of spraying gardenia plant by cumin oil as individual
treatment or in a mixture with clove or anise oil on the percentage of reduction
in the rotted buds caused by B.cinerea and the disease index of leaf spot
disease caused by A.alternata and P.langloissii under artificial inoculation
conditions showed that cumin oil at 2500 ppm mixed with clove oil at 5000
ppm concentration exhibited the heighst reduction percentage in the disease
incidence by the three tested fungi under artificial inoculation. Where the
treatment resulted in decreasing the incidence of gardenia bud rot disease
caused by B.cinerea from 58.3% in (check) treatment to 16.7% and also,
decreasing the incidence of gardenia leaf spot disease caused by A.alternata
and P.langloissii to 26.7% and 11.1%, respectively compared with the (check)
treatment 56.7 and 36.0%, respectively. Cumin oil at 2500 ppm mixed with
2674
J. Agric. Sci. Mansoura Univ., 33 (4), April, 2008
anise oil at 5000 ppm concentration treatment followed the previous mentioned
treatment. Meanwhile, cumin oil at 5000 ppm concentration was the lowest
one in this respect (Table 11).
Table (11): Effect of cumin oil individually or in mixture with anise or
clove oils on controlling bud rot caused by B. cinerea and
incidence of leaf spot disease caused by A. alternata and P.
langloissii.:
Plant
treatments
o ils
C. cyminum
C. cyminum
&
P. anisum
Cu. cyminum
&
S.aromaticum
(Check)
Mean
% infection of fungi
Conc. (ppm)
5000
2500
+
5000
2500
+
5000
---------------
LSD at 0.05 for:
Fungi (F)
Concentration (C)
:
:
A. alternata
2 8 .9
B. cinerea
5 0 .0
P.langloissii
2 3 .3
3 2 .2
4 1 .7
2 1 .0
2 6 .7
1 6 .7
1 1 .1
58.3
36.5
56.7
41.3
36.0
22.9
4.1
4.2
F×C:
7.2
2- Effect of cumin oil individually and in a mixture with anise or clove oil
on controlling bud rot and leaf spot diseases under natural infection
in greenhouse
The in vivo treatments by the same oils and by the same concentrations
were showed lower efficacy than those conducted under artificial inoculation
conditions in the greenhouse but in general, cumin oil at 2500 ppm mixed with
clove oil at 5000 ppm concentration exhibited the heighst percent of reduction
in the disease incidence of gardenia bud rot and leaf spot disease Where, the
treatment resulted in decreasing the incidence of gardenia bud rot disease
from 43.3% in (check) treatment to 23.4% by 46% efficacy and also,
decreasing the incidence of gardenia leaf spot disease from 41.7% in (check)
treatment to 25.0% by 40.0% efficacy .followed by cumin oil at 5000 ppm.
where, it reduced the incidence of gardenia leaf spot disease from 41.7% in
(check) treatment to 33.3% by 20.1% efficacy (Table 12). Kishnora et al
(2007), reported that very few studies have analyzed enough essential oils and
biological endpoints to determine whether there is a specificity for different
effects according to deferent oils or not . Clearly, it has been shown by Bakkali
et al. (2005 and 2006) that the essential oils presented a specificity in the
amplitude, but not in the mode of action, of the biological effects, i.e.
cytotoxicity, cytoplasmic mutant induction, gene induction and antigenotoxicity
effects. However, they did exhibit a specificity of the mode of action concerning
production of ROS, probably due to differences in their actual composition
corresponding to differences in compartmentation of the oxidative stress.
Hansen et al. (2006) concerning antigenotoxicity, the essential oils showed the
same protective activity. However the mode of action of protection differed, not
2675
Mosafa, M. A. et al.
according to the type of oil, but according to the mutagens, i.e. to the type of
lesions induced and thus, to the type of their enzymatic recognition and
processing lead to translational synthesis or late apoptosis/necrosis (Bakkali et
al., 2005 and 2006).
Table (12): Effect of cumin oil individually and in a mixture with anise or
clove oil on the percentage of reduction in rotted buds and
the disease index of leaf spot disease under natural infection
conditions in greenhouse.
Plant
treatments
Leaf spot
o ils
C. cyminum
C. cyminum
&
P. anisum
C. cyminum
&
S.aromaticum
(Check)
Mean
LSD at 0.05
Bud rot
% i n fe c ti o n
2 6 .7
% e ffi c a c y
3 8 .3
% i n fe c ti o n
3 3 .3
% e ffi c a c y
2 0 .1
2 9 .4
3 2 .3
4 1 .7
0 0 .0
2 3 .4
4 6 .0
2 5 .0
4 0 .0
4 3 .3
3 0 .7
2 .5
4 1 .7
3 5 .4
3 .1
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مقاومة امراض تبقعات األوراق واعفان البراعم على نبات الجاردينيا باستخدام بع ض
الزيوت النباتية الطبيعية.
محس ن احم د مت طفىج د عف ت عب د المجي د زا
نور الهدى عبد التواب رياضج
رجد عل ى حس ين ال
اعرجج و
ج قسم امراض النبات -كلية الزراعة -جامعة القا رة
جج قسم تعريف وحتر الفطريات – معهد بحوث أمراض النبات – مركز البحوث الزراعية -الجيزة
أظه ر تجارررا عجل مررنبجت ر جلجاارراتجل را ابلبرراجل ت رراجاججاجعمرراتجلرا ل جال ررا جل ج ر ل ج
ال ترتا اجت ج اعجتحافظاجل ربنةجخالبجتاس ج2006-2005جارااجأحاج ش جلا اجفط براجااجمراج
الحاج ش جرلساجفط باجتخال ا.جلخال تجلسعجارااجكبجفط جفيجل منبججاخاالفجكبجت جل رنءجل لجاايج
ل تمرررنابجتلررراجافاررر لتجل مرررنب.جاكالرررتجل ط بررراتججررراا لبا جسررربل با،جل ا لا بررراجل ا لاارررا،لاعجتررر ج
ل كالااسررجا ب جاجبساا اشررباجاللرلاسررباألجثرريجلركا ر جاكر ل لجفرريجل مررنب،جكترراجأظه ر تجثر جل مررنالتج
ل ط باجلر جماجل تخال اجلخاالفاجفيجقا اهاجل ت ضباج لىجل اجاجلررنلءجل تخال اج لجااراتجل را ابلبراج
حبثجلقا جل ط ججاا لبا جسبل باجفيجإ اجااج لج ل جقجبجا را جلرنثرا جاسرربجأ لرىجلسرجاجتئابراج
ر ررا جل جر ل جفرريجحررب جاخ ررفجكرربجتر جل طر ب جل ا لا برراجل ا لاارراجاجبساا اشررباجاللرلاسررباألجف ريج
إ رراجاه ج ررقا ل ججررا اجع جاكررا جل ط ر جل ا لا برراجل ا لاارراجثرراجلر لررىجفرريجشرراةجل رراجا.جأترراجل ط ر ج
كالااسجا ب جف شبجفيجإ اجااجالألجت جلرا ل جأاجل ج ل .ج
أظه ر تجارررا عجل تعااترراج ل ط برراتجل رراالثجل تخاج ر ةجالخرربجل تمترربججالسررطاج ش ر ب جنبارراج
ط باجأ جل نبتجل تسراخلفجتر جل كترا ج لراجل ا كبرنج500جررنءجفريجل تلبرا جألراجكرا جكافبراج ااجربطج
ل لتاجل طا يج ل ط براتجل تخاجر ة،ججبلتراجلحالرتجل نبراتجل تساخل راجتر جل بلسرا ،جل لملراع،جل ن ار ،ج
ل ع ل بجال بحا جل لسااألج لاجا كبنلتجاا لاحججب ج1000-750جرنءجفيجل تلبرا جل ت اجراجل االبراج
فيجث لجل شأ جالخالفجل ا كبنجل تاجطج كبجنبتججاخاالفجلاعجل ط .ج
كتاجأظه تجارا عجل تعااتاجاحتجظ افجل اجاججالسطاجل تماتلاججا نبتجل تساخلفجتر ج
ل كتا ججت اةجأاججمراجخلطراججرا نباتجل تساخل راجتر جل بلسرا جأاجل ع ل ربج لرىجل ارال يجأ جل تماتلراج
جخلرربطجتر جنبررتجل كتررا ججا كبررنج2500جرررنءجفرريجل تلبررا جانبررتجل ع ل رربججا كبررنج5000جرررنءجفرريج
ل تلبا جثاجلرفضبجفيجخ ضجحااثجل ت ض،جكتراج راحظجأ جك راءةجثر جل نبراتجفريجخ رضجحرااثج
ل ت ضجكالتجأ لىجاحتجظ افجل مااىجل لا باجتعا لراججك اءاهراجاحرتجظر افجل مرااىجل طجبمبراج
الخبجل اجا.ج
2680