Background and Objective: Bemisia tabaci (B. tabaci) is the most widespread insect pest of broad range of greenhouse and field crops. It is a serious threat to crop production due to direct damage. The aim of this study was to study the virulence of three entomopathogenic fungi against Whitefly, Bemisia tabaci. Materials and Methods: This study was carried out during 2 successive seasons (2015 and 2016). In this study trading compounds of entomopathogenic fungi, Bio magic (Metarhizium anisopliae), Bio power (Beauveria bassiana) and Bio catch (Verticillium lecanii) were used. Three concentrations were used (1×107, 1×108 and 1×109 spores mL1). Data were analyzed by one-way analysis of variance and followed by a least significant difference (LSD at 5%, p>0.05). Results: Under laboratory conditions V. lecanii, B. bassiana and M. anisopliae mortalities were happened after 3 days from treatment. The maximum percent of mortality (100%) was occurred after the 6th day from treatment with the 3rd concentration in all isolates. The 3rd concentration (1×109 spores mL1) was the highly toxic in V. lecanii, B. bassiana and M. anisopliae to the adult of B. tabaci compared with the other two concentrations. Under field conditions the 3rd concentration (1×109) also was the best concentration against whitefly after the third application in V. lecanii, B. bassiana and M. anisopliae. The percent of reduction was ranged between 52 and 100% in all concentrations. V. lecanii was more virulence than B. bassiana and M. anisopliae against B. tabaci. Conclusion: These results confirmed that V. lecanii, B. bassiana and M. anisopliae isolates are promising agents for whitefly control in the field.
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Whitefly, Bemisia tabaci is the greatest spread insect pest in greenhouse and field crops. It is the most dangerous to crop production due to direct damage1. Bemisia tabaci is split into 11 groups encompassing 24 species2. It can cause economic losses estimated in crops such as tomatoes, soybean, beans, cotton and leafy vegetables3,4.
Entomopathogenic fungi possess the unique ability to infect their host directly through the integument5.
The most promising fungi include V. lecanii, M. anisopliae and B. bassiana 6-17.
The V. lecanii, M. anisopliae and B. bassiana species have been used to control whiteflies and related insects in greenhouses in Europe, Canada and Egypt18-23.
The virulence of isolates of 3 entomopathogenic fungi (V. lecanii, M. anisopliae and B. bassiana) was determined on eggs, nymphs and adults of B. tabaci.
The cause of this increase is unknown but it may be due to the extended use of synthetic organic insecticides and subsequent augmented resistance to pesticides, changing climatic conditions24.
The entomopathogenic fungi B. bassiana has high activity against whitefly25. Blastospores and conidia can infect the host directly but mycelium needs to grow and from infectious propagates first26,27. The aim of this study was to study the virulence of 3 entomopathogenic fungi against whitefly, Bemisia tabaci.
MATERIALS AND METHODS
This study was carried out in greenhouse in El-Behira Governorate during season 2015-2016.
Entomopathogenic fungi: Bio magic (Metarhizium anisopliae), Bio power (Beauveria bassiana) and Bio catch (Verticillium lecanii).
Preparing of the concentrations: Three concentrations were used (1×107, 1×108 and 1×109 spores mL1) and add 0.5% tween 80. The spores were counted in the suspension using a haemocytometer (Swastik Scientific Company, India) blood cell counting chambers (Hirscmann 0.1×0.0025 mm2).
A haemocytometer is essentially a microscope slide bearing a small well of known depth. The base of which is marked with squares of known dimensions. During use the well is covered with a special coverslip (usually 0.4 mm thick).
Laboratory inoculation: Adults whitefly, B. tabaci were transferred to the laboratory from the greenhouse and put in Petri dishes with tomato leaf disk and incubated in 22±2°C and 85±5% RH. (Five adults/replicate) were used in all treatments. The entomopathogenic fungi were sprayed using a manual sprayer in a suspension containing (C1):1×107, (C2):1×108 and (C3):1×109 spores mL1, while sterilized water was sprayed to the leaves disks as blank control. The mortality of whitefly was observed daily for 7 day.
Greenhouse application: An area 30 m×10 m (= 300 m2) was divided into 4 parts each part was divided into 3 plots were treated with 3 concentrations from V. lecanii, M. anisopliae and B. bassiana and the other one as control treated by water. Every plot divided into three replicates. The suspensions were sprayed early in the morning and three times a day (one week interval). The live insects of B. tabaci per leaf/replicate were counted after all treatment.
The percent of reduction were calculated according to Henderson and Tilton28 as follows:
Tb and Ta are pre- and after-treatment counts, respectively.
Cb and Ca are untreated checks before and after treatment.
Statistical analysis: Data were analyzed by analysis of variance (one-ways classification ANOVA) and followed by a least significant difference (LSD at 5%, p>0.05)29.
Three concentrations of three isolates V. lecanii, M. anisopliae and B. bassiana were evaluated against B. tabaci under laboratory and greenhouse conditions.
Effect of V. lecanii, M. anisopliae and B. bassiana on B. tabaci under laboratory conditions: As mentioned in Table 1 there is no effect for V. lecanii, M. anisopliae and B. bassiana to B. tabaci after 3 days from treatment.
Mortalities are occurred in the 4th day. The percent of mortalities were increased gradually and reached to the maximum in the 7th day from treatment.
|Table 1:||Effect of V. lecanii, M. anisopliae and B. bassiana on B. tabaci under laboratory conditions at 22±2°C and 85±5% RH|
|(C1)1×107, (C2)1×108 and (C3)1×109 spores mL1, a-eDegree of significant from low to high, Mean±SD|
|Table 2:||Effect of V. lecanii, M. anisopliae and B. bassiana on B. tabaci in greenhouse during season 2015|
|a-cDegree of significant from low to high (±SD)|
|Table 3:||Effect of V. lecanii, M. anisopliae and B. bassiana on B. tabaci in greenhouse during season 2016|
|a-cDegree of significant from low to high, (±SD)|
With the all concentrations, the percent of mortalities were increased with increase of concentrations. The percent of mortalities ranged between 68.3-100, 56.2-90.5 and 62.7-97.3% with V. lecanii, M. anisopliae and B. bassiana, respectively, in the 7th day after treatment. Table 1 shows that there are slight differences between effect of V. lecanii, M. anisopliae and B. bassiana isolation to B. tabaci. This mean that V. lecanii isolation is more effective than M. anisopliae and B. bassiana. The percent of mortalities with all concentrations (C1, C2 and C3) of V. lecanii isolation were 68.3, 90.4 and 100%, respectively. The corresponding results with M. anisopliae and B. bassiana isolation were 56.2, 80.5 & 90.5, 67.8, 90.5 and 97.3, respectively.
Effect of V. lecanii, M. anisopliae and B. bassiana on B. tabaci in green house during season 2015: Table 2 obtained that the number of B. tabaci per leave was decreased compared with control after the second application. That the percent of reduction by V. lecanii M. anisopliae and B. bassiana after the third application were 57.3, 90.4 and 100, 50.2, 70.0 and 75.5 and 55.5, 79.5 and 90% with C1, C2 and C3, respectively.
The statistical analysis showed that there were no significant differences between the concentrations and control after the first application of all plots. After the 2nd application there were significant differences (p>0.05) between all concentrations and control. After the third application there were highly significant differences among all concentrations. The LSD was two, three after the third application. The statistical analysis confirmed that the third concentration (1×109) was the highly toxic compared the first and the second concentrations in all treatment.
Effect of V. lecanii, M. anisopliae and B. bassiana on B. tabaci in green house during season 2016: Table 3 show that after the second application the number B. tabaci leave were decreased compared with control. That the percent of reduction by V. lecanii M. anisopliae and B. bassiana after the third application were 55.2, 85.2 and 100, 50.0, 71.0 and 76.2 and 56.2, 78.3 and 92% with C1, C2 and C3, respectively.
The obtained data inferred that the entomopathogenic fungi V. lecanii, M. anisopliae and B. bassiana can be used as a promising agent in pest control and integrated pest management programs.
The effect of V. lecanii, M. anisopliae and B. bassiana may be depending on the type of host plant. While, the percent of mortality after 7th day by V. lecanii, M. anisopliae and B. bassiana ranged between 100, 72.2 and 92%, respectively28. This result compatible with Wraight et al.30, who found that both of B. bassiana and V. lecanii caused mortalities of up to 97 and 100% in Chilo partellus, respectively. Maniania31 reported that B. bassiana as an entomopathogenic fungi showed high effects on the aphid Aphis craccivora and the whitefly B. tabaci infesting cucumber. Zaki32 reported that V. lecanii caused higher virulence in the early stages of whitefly and reduced with older instars. Gindin et al.33, Abdel-Baky et al.34, Abdel-Raheem et al.35, Abdel-Raheem36, Abdel-Raheem et al.37, Abdel-Raheem et al.14 and Abdel-Raheem38 mentioned that entomopathogenic fungi caused good mortality to whitefly.
On the other hand, Sabbour and Abdel-Raheem39 described the control of the silver leaf whitefly Bemisia argentifolii on several plants (including cucurbits, broccoli, tomatoes and cotton) using B. bassiana; they observed only a few fungus-killed adults on the plants did not exceed 1%.
The results obtained that using of entomopathogenic fungi, V. lecanii, B. bassiana and M. anisopliae against B. tabaci as biological control agent are promising in the future. This study concludes that farmers have to use V. lecanii against B. tabaci in IPM program when first insect appear.
This study discovered the effect of Entomopathogenic fungi as biological control agents for insects. Verticillium lecanii, shows higher effect against Bemisia tabaci than Metarhizium anisopliae and Beauveria bassiana. Beauveria bassiana shows higher effect than Metarhizium anisopliae against Bemisia tabaci.
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