Amira, S. Mohammed

Ezzat, Gaballa Kassem Ahmed

Nagwa, M. M. El-khatteeb

International Journal of Horticultural Science

English

University of Debrecen Electronic Archive

International Journal of Horticultural Science 2013, 19 (1-2): 81-84
Agroinform Publishing House, Budapest, Printed in Hungary
ISSN 1585-0404
Introduction
Chemical control has been the main method of control used 
against T. absoluta in all tomato production regions in Argentina 
since 1970. Horticultural growers have tried to decrease its 
injure applying insecticides two times a week during a single 
cultivation period. Effective chemical control was difficult to 
achieve because of the mine-feeding behavior of larvae, lack of a 
threshold action, and deficient spraying technology (Marcela et. 
al., 2005).In addition to destruction of natural enemy populations 
(Campbell et al.,1991), build up of insecticide residues on tomato 
fruits (Walgen bach et al.,1991) and in the environment and 
fundamentally the rapid development of insecticide resistance 
(Siquira et al.,2000a and Siquira et al.,2000b).
Common insect pathogens used against insectpests 
according to Weinzierl, 1989 are Bacillus thuringiensis, 
B.popilliae, B.lentimorbus (bacterial pathogens) and 
Baeuvaria bassiana (fungal pathogen).
This investigation was carried out in the experimental 
farm at the faculty of Agricultural, Kafr El-Sheikh University 
during the two successive seasons 2011 and 2012 to study the 
following aims:
1. The effect of some uncommon bio- treatments used 
to control the tomato leaf miner; Tuta absoluta (Meyrick) 
infesting tomato plants.
2. Evaluating the percentages of infested tomato fruits by 
T. absoluta larvae in the tested treatments. 
Materials and methods
1. Feild studies
Field experiments were carried out at the experimental 
farm of the Faculty of Agriculture, Kafr El-Sheikh University 
during two successive seasons ; 2011 and 2012. One tomato 
variety namely Rosa was planted in the green house on April, 
21st and transplanted to the experimental field on May, 11 
and 15th of 2010 and 2011, respectively. The experimental 
area was 1/6 Feddan divided into seven plots. Each plote (80 
m²)was divided into three replicates. The normal agricultural 
practices of growing were done usual.
Six control treatments were used in the two tested seasons 
(four bio-treatments and two chemical insecticides), added 
to one untreated plot. Spraying treatments was started after 
one month from transplanting in the experimental field using 
cp
3
 Knapsack. Spraying was applied twice a week until the 
tomato leaflets dryness.
Leaflet samples were taken randomly after twenty four 
hours from applying treatments.A sample was consisted of 
Some microbial treatments against the tomato leaf miner; 
Tuta absoluta (Merick) under natural field conditions
Amira S. Mohammed and *Nagwa M. M. El-Khatteeb **Ezzat Ahmed
Entomology Department, *Botany Department Faculty of Agriculture, Kaferelsheikh University. Egypt. 
**University of Debrecen Centre of Agricultural Science and Engineering Institute for Research and 
Development, Hungary
Abstract: In an attempt to find more effective methods and safety to control the tomato leaf miner, Tuta absoluta, this research was conducted 
to study the efficacy of the four bio-treatments ; two bacterial isolates (Bacillus subtilis. and Pseudomonas Fluorescence), one fungal isolate 
(Trichoderma viride) and Spinosad as a microbial insecticide compared with the most common insecticides used against the dipterous and 
lepidopterous leaf miners in Egypt; Profenofos and Aphox. In the two successive seasons; 2011 and 2012, the fungal isolate Trichoderma 
viride achieved the lowest infestation(numbers of infested leaflets, mines, infestation percentages) which were 167, 195, and 21.9% resp.in 
season 2011 and 198, 222 and 26.2% resp. in season 2012. While the highest infestation was occurred in case of using the insecticides, Aphox 
(423, 559 and 55.9% resp.& 384, 839 and 72.2%. and Prohpenofos (436, 562 and 57.7% &391, 534 and 51.7% resp.).during 2011 and 2012 
respectively. The other treatments occupied the intermediate levels.
On the other hand, the yield of tomato fruits was increased by 75% over the check in case of plants treated with Trichoderma viride isolate, 
but the pesticide treatment given weak or no impact in increasing tomato crop.
In season 2012, the infested tomato fruits percentages with T. absoluta larvae and Rhizoctonia soil rot disease(that Frequently observed 
incidentally) were recorded the minimum levels obtained by spraying Trichoderma viride isolate(21.38 and 10.32% resp.). In contrast, the 
maximum levels were recorded in Prophenofos (44.87 and22.78% resp.) and Aphox treatments(47.22 and 27.78% resp.).
Keywords: tomato
82 Amira S. Mohammed and Nagwa M. M. El-khatteeb Ezzat Ahmed
108 leaflets/3 replicates for each treatment. Samples were 
transferred to the laboratory to examine numbers of infested 
leaflets, mines and estimating the infestation percentages.
Fruit samples were collected weekly for one month.Three 
plants were selected randomly to pick up its fruits for each 
treatment. Mean tomato fruit yield, infested tomato fruits by 
Tuta absoluta larvae and infested tomato fruits by the tomato 
leafminer larvae& fungi were evaluated.
2. Tested materials.
2.1. Bio-treatments:
2.1.1. Microbial isolations:
2.1.1. (a). Bacillus subtilis.
2.1.1. (b). Pseudomonas Fluorescence
2.1.1. (c). Trichoderma viride
2.1.2.Bacterial insecticide.
2.1.2. (a). Spinosad:
Spinosad dust (0.125%WP) Dow AgroSciences product 
is a mixture of the two most active naturally occurring 
metabolites of the soil bacterium Saccharopolyspora 
spinosad. Spinosad is a mixture of about 85% Spinosyn 
A and 15% Spinosyn D.,structurally, these compounds 
are macrolides and contain an unique tetracyclic ring 
system to which two different sugars are attached(Abdel-
Mageed,2008).
Spinosyn A, Empirical Formula C41H65NO10;MW 
731.98.
Spinosyn D, Empirical Formola C42H67NO10; MW 
746.00. 
2.3. Chemical insecticides:
2.3.1. Profenofose:
Common name is Selecron (72% EC).It is an 
organophosphorus compound applied at rate of 100cm/100L 
water.
2.3.2. Aphox:
The common name is Pirimicarb(50%DG). It is a 
carbamate compound applied at rate of 250 gm/100L water.
The insecticides used against all leaf miners in Egypt are 
also used to control the white flies and aphids which have 
systemic effect. 
3. Laboratory studies:
3.1. Preparation of microbial inocula: 
The microorganisms were recultured on suitable liquid 
media, potato dextrose broth (PDB) for fungi or nutrient broth 
(NB) for bacteria in flasks of 500 ml capacity containing 200 
ml medium. In case of fungus, cultures were incubated at 28o 
C for 10 days. Spore suspension of Trichoderma isolate was 
sieved through double cheesecloth and hand homogenized 
and counts of spores were adjusted to 106spore/ml using 
hemocytometer. In cases of bacterial isolates, cultures were 
incubated at 28o C for 5 days in shaking incubator (140 r/
min.) and cell suspension was adjusted to 108 cell/ml. Bacillus 
subtilis, Pseudomonas fluorescence and Trichoderma viride 
isolated by El-Khateeb (2004) were kindly obtained.
3.2. Procedure of application:
Cultures were amended with calculated aliquots of an 
adhesive surfactant (New-Film, 1265 registered by ministry 
of Agric., Egypt) as recommended (30 ml/100 L) and hand 
homogenized before fine spraying onto the upper and the 
lower leaf surfaces of plant cotyledons until runoff. Plants, 
sprayed with sterilized tap water (likely amended with the 
adhesive surfactant), served as check treatment (control). 
3.2. Identify the pathogens.
During the examination of tomato leaflets samples picked 
from plots treated by Trichoderma viride, some larvae were 
found showing symptoms of microbial disease. Larvae had 
soft bodies, static, cuticle color was close of brown. The 
pathogen was isolated from the diseased larvae to be defined. 
Results and discussion
1. The effect of some bio- treatments used to 
control the tomato leaf miner; Tuta absoluta 
(Meyrick) infesting tomato plants. 
Table (1) and figure (1) showed the number of infested 
tomato leaflets, the number of mines and the infestation 
percentages in the two successive seasons; 2011and 2012 
according to the four bio- treatments tree isolations and one 
bioinsecticide) and two chemical insecticides. In the first 
season, the Tricoderma sp. isolate gave the least infestation 
(167, 195, and 21.9% resp.) followed by Spinosad (233, 
301 and30.8%), the Bacillus subtilis isolate (246, 338 and 
32.9% resp.), the Pseudomonas Fluorescence isolate (334, 
475 and 44.2% resp.). In the latest ranking, Aphox (423, 559 
and 55.9% resp.) and Prohpenofos(436, 562 and 57.7%). 
The nearby trend was recorded in the second season 2012. 
Since the lowest infestation level related with Tricoderma 
viride treatment (198, 222 and 26.2% resp.).The middle 
infestation levels related with Bacillus subtilis,Spinosad.and 
Pseudomonas Fluorescence treatments (261, 351 and 34.5% 
& 270, 383 and 35.7% & 321, 466 and 42.4% resp.).While 
Aphox and Prophenofos recorded the highest infestation 
levels (384, 839 and 72.2% &391, 534 and 51.7% resp.).
Generally, the chemical insecticides Aphox and 
prophenofos recorded greater infestation than microbial 
treatments (Tricoderma viride,Bacillus subtilis,Pseudomonas 
Fluorescence) and Spinosad. While the greatest values were 
obtained in check during the two seasons 2011 and 2012 
(614, 889 and 81.2% & 546, 839 and 72.2% resp.) Statically 
analysis showed significant differences among the tested 
treatments in their efficiency of T. absoluta control.
Figure (1) showed one of the diseased larvae that was 
obtained during the examination of samples. Leaflet samples 
taken from plants treated with Trichoderma viride isolate. 
83Some microbial treatments against the tomato leaf miner; Tuta absoluta (Merick) under natural field conditions
The pathogen was isolated and defined by Microbiology 
Laboratory,Botany Department in Faculty of agriculture 
at Kafer elsheikh University. The pathogen was defined as 
Trichoderma viride.
These findings agreed with Lietti et. al., 2005 they 
reported that some populations of T. absoluta have developed 
resistance to organophosphate and pyrethroid pesticides. 
Gonzalez-Cabrera et. al., 2011 mentioned that newer 
compounds such as spinosad and Bacillus thuringiensis 
have demonstrated some efficacy in controlling European 
outbreaks of this moth.
2. Evaluating the percentages of infested 
tomato fruits by T. absoluta larvae in the tested 
treatments.
Data presented in Table (2) indicated that the maximum 
yield of tomato fruits during four weeks was took place in 
Tricoderma viride treatment (126 fruits/12 plants.) which 
increased by 75% over the check. While the minimum yield 
was occurred in Aphox treatment (72 fruits/12 plants)that 
equaled with check. The tomato fruits yield resulted from 
Bacillus subtilis, Spinosad and Pseudomonas Fluorescence 
treatments were occupied intermediate values (100, 97,and 
91 fruits/12 plants resp.).
Concerning the infested tomato fruits percentages with T. 
absoluta larvae in season 2011 were arranged in ascending 
order as follows; 21.38, 37.00, 37.36, 38.14, 44.87, 47.22 and 
55.56% in Tricoderma viride, Bacillus sutilis, Pseudomonas 
Fluorescence. Spinosad Prophenofos, Aphox treatments and 
finally check respectively.
During the examination of infested fruits by T. absoluta 
larvae, Found that a large number of them infected with 
Rhizoctonia solani fungus that causes Rhizoctonia soil rot 
disease. Symptoms are alternating light-and dark-colored 
zonate bands on fruit surfaces (figs.3 and 4). The infected 
tomato fruits must had a hole of tomato leaf miner larva 
(fig.2).The infected tomato fruits with fungus ranged between 
41.94 and 60.00% from the total of infested tomato fruits 
with T. absoluta larvae according to the different treatments. 
The highest infestation percentage was obtained by spraying 
Table (1): The effect of the four bio- treatments compared to chemical insecticides against T. absoluta during 2011 and 2012 seasons.
Seasons 2011A 2012B
Treatments Inf. leaflets mins. No. Infestation% Inf. leaflets mins. No. Infestation%
Bacillus subtilis 246d 338d 32.9e 261d 351d 34.5d
Spinosad 233d 301d 30.8e 270d 383d 35.7d
Pseudomonas Fluorescence 334c 475c 44.2d 321c 466c 42.2c
Tricoderma viride 167e 195e 21.9f 198e 222e 26.2e
Prophenofos 436b 534b 51.7c 391b 562b 51.7b
Aphox 423b 559b 55.9b 384b 516b 50.8b
Control 614a 889a 81.2a 546a 839a 72.2a
The values were an average of three determinations.
Values followed by the same letter in column are not significantly different at p < 0.01.
a, b, c, d, e, f and g: comparison of means by treatments.
Fig.(1): Infected larva after Trichoderma viride treatment.
Table (2): The percentages of yield and infested tomato fruits under tested treatments in 2012 season.
Treatments. yield⁄12 plants
Average 
yield⁄plant
%yield 
increasing
Infested fruits by
Larvae Larvae + fungi
Total % Total %
Bacillus subtilis 100b 8.33b 38.89b 37b 37.00e 18d 18f
Spinosad 97b 8.08bc 34.72c 37b 38.14d 21b 21.65de
Pseudomonas Fluorescence 91c 7.58c 26.39d 34c 37.36de 19cd 20.88e
Tricoderma viride 126a 10.50a 75.00a 31d 21.38f 13e 10.32g
Prophenofos 78d 6.5d 8.33e 35c 44.87c 18d 22.78c
Aphox 72e 6e 0.00f 34c 47.22b 20bc 27.78b
Control 72e 6e 0.00f 40a 55.56a 24a 33.33a
The values were an average of three determinations.
Values followed by the same letter in column are not significantly different at p < 0.01.
a, b, c, d, e, f and g: comparison of means by treatments.
84 Amira S. Mohammed and Nagwa M. M. El-khatteeb Ezzat Ahmed
Aphox(58.82%) then the control (60.00%), while the lowest 
infestation percentage was achieved by using Tricoderma 
viride isolation which was 41.94% (Table 2 and Fig.2).
The previously data can be interpreted that the tomato 
leaf miner larvae speed up the entry of Rhizoctonia solani 
fungus. So, the real danger of Tuta absoluta lies in its impact 
on quantity and quality of the tomato crop. 
Fungi of the genus Trichoderma are important biocontrol 
agents (BCAs) of several soil borne phytopathogens 
(Benítez et al. 2004). The biocontrol by fungal species of 
the Trichoderma genre, of root and crown rot caused by R. 
solani, are being used as an alternative to chemical fungicides 
(Papavizas, 1985; Limon et al. 1999; Rey et al. 2001). 
Statistical analysis for yield increasing percentages 
showed significant differences among all experimented 
treatments. While the infested tomato fruits by T. absoluta 
larva and by T. absoluta larva with fungi revealed that 
Trichoderma viride differ highly significant than the 
others. 
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Infested fruits by larvae
Series1
Series1
6 5
4 3
2 1
50
45
40
35
30
25
15
20
10
5
0
Series2
Series3
Series3
Series4
Infested fruits by larvae&fungi
%
 infested tom
ato fruits
Fig. (2): The percentages of infested tomato fruits by T. absoluta larvae and 
T. absoluta larvae and fungi in the tested treatments
Fig. (3 and 4): Tomato fruits infected with Rhizoctonia soil rot disease.


Some microbial treatments against the tomato leaf miner; Tuta absoluta (Merick) under natural field conditions

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Some microbial treatments against the tomato leaf miner; Tuta absoluta (Merick) under natural field conditions

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