Efficiency of essential oils emulsions against Whitefly Bemisia tabaci (GENN.) infesting potato plants under field conditions In Egypt

doi:10.21203/rs.3.rs-2859165/v1

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Efficiency of essential oils emulsions against Whitefly Bemisia tabaci (GENN.) infesting potato plants under field conditions In Egypt

https://doi.org/10.21203/rs.3.rs-2859165/v1

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The whitefly, Bemisia tabaci, nymphs, and adults sucking sap Excessive use of pesticides caused pollution of the environment and the death of beneficial insects, it is one of the most damaging pests of Potato, Solanum tuberosum, so it was necessary to search for more safe alternatives. An experiment was carried out during seasons 2021 and 2022 under field conditions in Egypt. The objective of this study aimed to use peppermint and eucalyptus essential oils and prepare coarse emulsions and nanoemulsions (CE and NE) of both peppermint and eucalyptus essential oils.

The results reported that the numbers of nymphs per plant before treatment during season 2022 were greater than those in season 2021, which may be due to increasing temperature and decreasing precipitation, specific humidity, and wind speed. On the other hand, essential oil's nanoemulsion (NE) was more effective in controlling B. tabaci. Generally, the toxicity decreased with time after spraying. The second spray was more effective than the first spray. P (CE, NE) revealed the most effectiveness, followed by E (CE, NE) during both seasons. During season 2021, the NEs were slightly more effective than the CEs. During season 2022, recorded no significant difference between CEs and NEs. Also, the toxicity of the tested emulsions was highly effective during season 2021 compared with 2022 due to decreasing temperatures in December 2021 compared to December 2022. The study of the growth component deduced that the parameters of potato plants after being treated with tested emulsions showed that both peppermint coarse emulsion PCE and eucalyptus coarse emulsion ECE achieved slightly decreased portion levels. ECE increased total plant carbohydrates. However, our treatments did not affect the phenolic compounds of potato leaf plants, although ENE caused an increase in phenolic compounds. All treatments decreased the nitrogen plants' contents. Furthermore, PCE, PNE, and ENE increase the potassium content. All treatments increase the activity of peroxidase (POX) compared with untreated plants. The formulation obtained here might be an interesting alternative for integrated pest management of B. tabaci nymphs.

Bemisia tabaci

potato plants

essential oils

nanoemulsion

weather conditions

Potato, Solanum tuberosum, is one of the most important root and tuber crops for food. It ranks fifth in the world among the most important staple foods (Orlando et al., 2020). It is third in terms of consumption and fourth in terms of production of tuberous crops (Bahar et al., 2021). In 2017, Egypt's potato production was around 4.33 million tons from a harvested area of approximately 163,939 hectares (FAO 2020). Egyptian potatoes are locally consumed, processed, or exported abroad. For exportation, potato is the largest horticultural export, with about 561.4 thousand tons of potato exported in 2020 (El-Din et al., 2022). Potatoes, like all other crops, are infested by a variety of insect pests at various stages of growth, which impacts potato agriculture and functions as a limiting factor in the economic cultivation of this crop. These pests are classified into storage pests, soil pests, sap feeders, and foliage feeders, of which sap feeders (aphids, whiteflies, thrips, and armyworms) are the most destructive pests, causing damage to the foliage of the crop at the vegetative stage (Bojan, 2021).

Essential oils (EOs) have been known for insecticidal activity against a large variety of insects, herbivores, and microorganisms, and some are active in attracting pollinators (Argyropoulou et al., 2022; Meena et al., 2022). Previous studies have shown their contact toxicity, repellency, pathogenicity, and growth regulation (Li et al., 2022; Singh and Pandey, 2021; Sahu and Singh, 2022). The important challenges for applying EOs in agriculture are attributed to their poor solubility in water, rapid environmental degradation, and limited physical stability (Abdelaal et al., 2021).

and extending potential applications have been widely considered. Toxicity of EO-based CE sand NEs was tested on several insects of agricultural and medical interest such as sweet potato whitefly (Bolandnazar et al. 2020), aphids (Heydari et al. 2020), Cotton bollworms (Moustafa et al. 2015), stored-product beetles (Adak et al. 2020; Hossain et al. 2019),

The objective of this study is to identify the main constituent compounds of peppermint and eucalyptus essential oils, prepare CE and NE of both peppermint and eucalyptus essential oils, and evaluate their efficiency against the whitefly, B. tabaci, in potato cultivars during seasons 2021 and 2022. Also, study the role of weather conditions on the emulsion's toxicity against the tested insect. Finally, study the effect of these emulsions on the chemical components of potato leaves.

Tested materials

This trial was designed to evaluate the efficacy of two forms of tested emulsions; peppermint course emulsion (PCE), peppermint nanoemulsion (PNE), eucalyptus course emulsion (ECE) and eucalyptus nanoemulsion (ENE). Essential oils were purchased from El-Gomhoria chemical company, Cairo, Egypt. The oils were kept at 4°C until experimentation. Oils emulsions efficacy was evaluated compared with Imidacloprid as a standard pesticide on the population of Bemisia tabaci. Suncloprid ® Imidacloprid 35% S.C was obtained from El-Nasr Company for Intermediate Chemical, Giza, Egypt, it was applicated at the rate of 75 gm/ 100 L water.

GC-MS analysis

The chemical analysis of Peppermint and eucalyptus essential oils were carried out by Gas Chromatography-Mass Spectroscopy (Trance GC ULRA, Thermo) coupled with ISQ Single Quadruple mass spectroscopy with direct capillary column TG-5MS (30 m, 0.23 mm, 0.25 film thickness). Initial temperature 45^oC for 2 minutes, then programmed to 165^oC at 3^oC min-1 for 10 minutes. The temperature was then increased to 280 at 15oC min-1 and kept for 10 minutes before being increased to 350 ^oC till the conclusion. Helium was used as the carrier gas, with a constant flow rate of 1 ml min-1. The scan mode (30–500 aum /0.2 Sec) was used for the sector mass analysis. Spectra were obtained using the EI mod with an ionization energy of 70 eV. By comparing the mass spectra of each peak to those in the library, the components were identified.

Emulsion preparation

Oil-in-water coarse emulsions were prepared by stirring (5% essential oil, to 10%, tween 80 and 85%, v/v) distilled water at approximately 4000 rpm for 20 min. Nanoemulsions were prepared by ultrasonication of the coarse emulsion at 20Mz for 30 min (Sugumar et al. 2013). The emulsions were stored at 4ᴼC. Nanoemulsions of the test oils were characterized by measuring the particle size, and polydispersity index (PDI) by photon correlation spectroscopy using Zetasizer Nano ZS (Malvern Instruments, UK), then kept for further experiments.

Experimental area

A field experiment was carried out during the subsequent growing potato season 2021, and 2022 at the Plant Protection Research Station (PPRS) at Qaha, Qalubiya Governorate, Plant Protection Research Institute Egypt, (PPRS) located at a distance of 29.1km (44 mins) of Cairo. Latitude:30 1719.14 N Longitude: 31 1151.33 E. It is located in the rich farmland of the southern part of the Nile Delta and is characterized by well-irrigated canals leading off the Delta Barrage. The experimental area was about 396 m² and was cultivated with potato, Solanum tuberosum (Spunta) on the 18th of September in both seasons. Treatments were arranged in a split-plot-plot system in a randomized complete blocks design (RCPD) divided into 6 treatments (each treatment replicated 3 times) for 6 treatments each plot was 21 m²: (1) the control treatment ( sprayed with water ); (2) the plots treated with the bio-insecticide PCE (3) the plots treated with the PNE; (4) the plots treated with ECE (5) the plots treated with ENE (6) and the plots treated with Imidacloprid (positive control)considered as the reference at rate 75gm/ 100 L water. Each plot covers an area of 21 m2 (7 m x3m) and contains 33 plants (3 rows of 11 plants were cultivated with a spacing of 0.60 m. The treated plots were separated from each other by untreated rows as a barrier to prevent overlapping treatments.

Sampling methods

Once weekly visit, 45 days after planting and continued till harvesting, each visit picked up the main samples of 100 potato leaves were randomly selected and transferred into paper bags for examination by the stereomicroscope. Inspection of plant leaves was carried out before spraying (pretreatment) and after 3, 7and 10 days after application respectively. The reduction percentage of nymphs/each treatment was calculated according to Henderson's and Tilton's formula (Henderson and Tilton 1955).

Relationships between some phytochemical components of potato leaves and density of B. tabaci pest.

We determine certain phytochemical components in the leaves of potato to explain the relationship between growth leaves components (nitrogen, phosphorus, and potassium) and infestation with the studied pest during the plant growth period. The analysis of leave samples was conducted in the Department of Insect Physiological, Plant Protection Research Institute, Agricultural Research Centre.

Estimation of total soluble proteins

Quantitative valuation of protein was performed by the (Bradford, 1976) method. Absorbance was measured at 595 nm.

Estimation of total carbohydrate

Preliminary hydrolysis to convert polysaccharides into monosaccharides was performed by using 2.5N HCl. The total carbohydrate content was estimated by the phenol-sulfuric acid method (Dubois et al. 1956).

Estimation of total phenolic contents

The fresh leaves were extracted as IAA extraction, the 0.5 ml extract, 0.5 ml carbonate, and distilled water were added to each tube, shaken, and allowed to stand for 60 minutes. The optimal density was determined at 725 nm using a spectrophotometer (Singleton and Rossi, I965; Diaz and Martin GC 1972).

Estimation of peroxidase enzyme activity

Peroxidase activity was established at 26°C with a spectrometer (PD-303UV) at 470 nm using guaiacol and H₂O₂ (Ponce et al. 2004).

Statistical Analyses

Data were statistically analyzed by the significance of mean differences between treatments and controls were compared statistically using twice analysis design. Firstly, with one-way complete block and least significant difference (LSD₀₅) significant differences (P < 0.05). A second analysis by variance (split-split-plot ANOVA) and at the 0.05% probability level, Main, sub, and sub-subplot means separation was tested with Tukey’s multiple range tests at a 95% confidence level via Co-Stat software. The data were calculated by date, and the data were analyzed for each year separately.

Chemical composition of peppermint and eucalyptus essential oils

The essential oils of both peppermint and eucalyptus components were identified by GC-MS. The main constituents of peppermint and eucalyptus oils are shown in Table (1) & Figer (1); peppermint oil has 11 components that were characterized, representing 99.00% of the essential oil. A Large number of oxygenated monoterpenes and sesquiterpene derivatives. The main components discovered in this fraction were menthol (35.90%), menthone (32.53%), 1, 8-cineole (11.26%), caryophyllene (4.29%), and (sabinene) or sabonine (3.25%). Amongst them, the significant two components of Peppermint are menthol and menthone. On the other hand, eucalyptus oil has 32 components described and represents 98.87% of the essential oil. The main components were 1, 8-cineole (27.78%) followed by alpha-terpinene (19.78%), 1-p-menthol (18.37%), monoelaidin (13.39%), and çterpinene (5.09%).

Nano-emulsions characterization

Data of all these Nano-emulsions were successful in their preparation in the nanometric size range. In Table (2) & Figure (2), the PNE droplet size distribution was 114.5 nm with PDI 0.27. The ENE droplet size distribution was 145.8 nm with PDI 0.69.

Weather data

The seasons 2021 and 2022 are at Qaha, Qalubiya Governorate, Egypt. Latitude: 30 17 19.14 N Longitude: 31 11 51.33 E differed dramatically in weather-related abiotic factors, particularly in the amount of daily average precipitation (Fig. 3). From 18 September to 31 December, the primary month of field trials, the daily average temperature was 12.16 ◦C in 2021 and 12.01°C in 2021. December 2022 was much warmer than December 2021; the daily average precipitation was 0.38 cm in 2021 and 0.13 cm in 2022. As well, the average specific humidity is 9.05 g/kg, compared to 8.96 (g/kg). The daily average wind speed in 2021 was 2.53 m/s, compared with 2.35 m/s in 2022. All the weather data were collected by the Central Laboratory of Agricultural Climate, Agriculture Research Center, Dokki, Giza, Egypt.

Efficacy of tested emulsions on reduction % of B. tabaci (nymph) infesting potato plants

The average number of B. tabaci nymphs infesting potato leaves before the application were 15.00 to 18.66 nymphs per plant during the 2021 summer season experiment, while the average number increased from 23.16 to 26.66 nymphs per plant during the 2022 summer season experiment, respectively. Differences in whitefly infestation levels were detected (P < 0.05) among treatments and Imidacloprid as a positive control for the first and second post-treatment observations across the three post-treatment observations in seasons 2021 and 2022. The numbers of nymphs per plant and reduction percentages of infesting potato plants as influenced by 5% P (CE, NE) and 5% E (CE, NE), which were applied to potato plants in two consecutive sprays during season 2021, were tabulated in Tables (3 and 5). During the whole duration of the study, we found a significant difference between all treatments compared with the control. The second spray was more effective than the first spray. After the first spray, PNE was slightly more effective than PCE, where the reduction percentage was (83.13, 67.12%) and (82.09, 60.65%) after 3 and 10 days from application, respectively. In the same trend, after the second spray, the ENE was slightly more effective than the ECE, the reduction % was (73.26, 72.72%) and (67.00, and 61.90%) after 3 and 10 days from application, respectively. During season 2022, the field experiment revealed similar data to season 2021 field experiment Tables (4 and 5) & (Fig. 3). All treatments showed significant differences compared with untreated plants. Generally, the toxicity decreased with increasing time after spraying. Also, the second spray was more effective than the first spray. P (CE, NE) revealed more effectiveness against B. tabaci nymphs, followed by E (CE, NE). There is no significant difference between PCE and PNE after the first spray. The mean reduction% in case P (CE, NE) was 79.88 and 76.96% after 3 days from spray; these results reached 66.17, and 65.05% after 10 days from spray compared with 87.22 and 73.85% with Imidacloprid. E (CE, NE) produced the least reduction% due to the first spray (73.36, 68.78%) and (77.85, 69.79%) after the second spray. Generally, during season 2022, there was no significant difference between CEs and NEs.

Effects of tested emulsions on phytochemical components of potato leaves.

The growth components parameters of potato plants after being treated with tested emulsions were tested in Table (6) both CEs showed a slightly decreased in protein levels. Its level was 9.73 (mg/ g fresh weight) compared with 11.31(mg/ g fresh weight) in untreated potato leaves. The ECE increased total plant carbohydrates from 47.53mg/g within control to 57.10 mg/g followed by 52.2 and 50.93 mg/g with PNE and ENE, respectively. Phenolic compounds of potato plants do not affect our treatment expected ENE which caused increases in phenolic compounds 2.06 mg/ g compared with1.92 mg/ g with control. All treatments decreased nitrogen plants' contents. On the contrary, all tested emulsions increase phosphorus and potassium contents.

Effects of tested emulsions peroxidase enzyme

Determination of the potato plant antioxidant enzyme peroxidase after being treated with essential oils CE and NE under studied tabulated in Table (6). All tested emulsions increase peroxidase (POX) activity compared with untreated plants. The ENE most affected in the enzyme activities was 621.00 (∆ O.D. /min/g) followed by PCE, PNE, and ECE 521.66, 521.66, and 515.33(∆ O.D. /min/g), respectively. These materials increase the activity of the Peroxidase enzyme compared to control, where this enzyme great role play in oxidation and reduction processed inter plants.

The goal of the current study evaluates the efficacy of the CEs and NEs of peppermint and eucalyptus oils as a potential botanical insecticide to control whitefly, Bemisia tabaci pest in Qaha, Qalubiya Governorate, Egypt. The efficacy of tested emulsions in this study was slightly less than that of the conventional synthetic insecticide, Imidacloprid, in managing the B. tabaci on potato fields. I used peppermint and eucalyptus oil used peppermint and Eucalyptus oil as a bio-pesticide to control insects to protect the environment from pollution, use these materials because of the two terpenoids (α terpinene, C terpinene, menthol, and menthene) components. The present result was in concordance with the previous studies (Heydari et al. 2020; Rajkumar et al. 2020; Beigi et al. 2018; Buleandra et al. 2016). The menthol has cholinesterase inhibitory effects and is reactive to nicotinic, 5-hydroxytryptamine (HT3), GABAA, glycine, and receptors (Kennedy,2019). Also, the major contents of eucalyptus oil were 1-p-menthol,1, 8-cineole, and alpha-terpinene. Many researchers recorded that the primary component in eucalyptus was 1, 8-cineole (eucalyptol) (Chaudhari et al. 2021; Chauhan et al. 2018; Bett et al. 2016). They inhibited acetylcholinesterase and binding to gamma-aminobutyric acid (GABA_A), nicotinic, and muscarinic receptors (Kennedy,2014). I could attribute the components and quantity of essential oil in the same plant species to the differences in processing conditions, such as harvesting time, geographical region, seasonal factors, and method of extraction (Pang et al. 2020). Although, peppermint and eucalyptus oil showed bio-pesticide effects to control Whitefly (Aroiee et al. 2005; Bolandnazar et al. 2018). but, the difficulty in applying essential oils to control insects on a field scale and under environmental conditions required the investigation into new formulations through nanotechnology. So, we prepared our successful nanoemulsions in the manometric size ranges PNE and ENE were 114.5, and 145.8 nm. Smaller droplet sizes lead to NEs with improved more stable, which is a significant factor for many applications. Many researchers found NEs having a particle size between 10 and 00 nm showed beneficial imputes, such as solubility and permeability (Almadiy et al. 2022; Abdelaal et al. 2021). The PDI value of PNE showed good physical stability of the NE, because of the reduced Ostwald ripening (Hashem et al., 2020). Whereas, a PDI value of ENE above 0.30 shows heterogeneity of the NE (Nenaah et al. 2022). To whitefly manages, a less amount of (EO) NE may be required than bulk EO. Downsizing the oil particles may have allowed it to come into contact with insects, as opposed to EO, which showed its toxic effect. This was consistent with the findings of (Heydari et al. 2020; Barzegar et al. 2018). However, the smallest size of the NE formulation of PNE obtained by (Shaker et al. 2022) was about 66 nm, and ENE obtained by (Mohammadi et al. 2020) was about 103.9 nm, which is like the present research. Smaller particle size improves the NE penetration through the insect cuticle and plays a critical role in insecticidal activity. Low particle sizes of NEs increase penetration and uptake of EO into the insect tissues. This may improve the biological activity of NEs compared to CEs (Mustafa and Hussein 2020; Adak et al., 2020).

Despite many essential oils estimated as insecticides, only a few researchers have focused on vegetable crops in the field. There are very few studies on crop systems that have investigated both the effectiveness and the impact of insecticide treatments on the crop in the field. Also, most articles discuss in toxicity of certain essential oils and nano-formulations. The effectiveness of both NE and CE in this study was slightly less than that of the synthetic insecticide, Imidacloprid in managing the major whitefly population in potato fields. Similarly positive results were regarded with the NE of Lippia multiflora Mold. against the reduction of the cabbage pest. The NE was very effective compared with the synthetic insecticide lambda-cyhalothrin (Tia et al. 2021). The NE of Mentha piperita (EO) except high contact toxicity on the cotton aphid in the laboratory condition (Heydari et al. 2020).

The efficacy of tested emulsions heavily depended on the weather, as seen in the contrast between the 2021 and 2022 field trials. The density of the B. tabaci population during seasons 2021 and 2022, respectively, was in harmony with the weather parameters, i.e.; temperature, precipitation, Specific Humidity, and wind speed. This was clearly in the numbers of nymph per plant before treatment during season 2022 was greater than those in season 2021 may be because of increasing temperature and decreasing precipitation, Specific Humidity, and Wind Speed. The weather parameters affect the population dynamics of B. tabaci on tomato crop temperature had a negative correlation, while relative humidity had a positive correlation, but wind speed had a non-significant positive correlation (Jha and Kumar 2017). Our data showed that the toxicity of tested P (CE, NE) and E (CE, NE) was highly efficacy due to decreasing temperatures in December 2021, correlated with December 2022. Thymol and carvacrol exhibited significantly higher efficacy at the lower temperature of 15°C than at 30°C (Pavela and Sedlák 2018). The toxicity of tested emulsions showed decreased after 10 days of the second spray may be because of rainfall. The emulsions were highly susceptible to being washed away by rainfall (Johnston et al. 2022).

The results showed that sprayed with both tested NEs doesn’t affect negatively total protein levels in potato leaves. Our outcome shows we can control insects in the field without affecting plant protein contents conversely conventional insecticides decrease the total protein accordingly (Siddiqui and Ahmed 2002 and 2006). Also, the result refers to the positive effect of all tested emulations on carbohydrates in potato plants. Pandey et al. 2020 found there were no significant differences between carbohydrate content in untreated tomato plants and others treated with PNE (Pandey et al. 2020).

All treatments did not affect total phenol except ENE has a positive effect which recorded maximum total phenolic contents of 2.06 mg/g and therefore activates phenolic compounds production. increases of phenolic compounds induced in plants are direct toxicity to insects or produce toxic secondary metabolites and activate the defensive enzymes Bhonwong et al. 2009). our data agrees with (Pandey et al. 2020) who found no significant difference between plant PNE and control. Plants use phenolic compounds for a variety of purposes, including growth and development, cell wall thickening, hormone synthesis, pigmentation, reproduction, stress resistance, osmoregulation, UV protection, anti-herbivore and antibacterial activities (Dixon, 2001). also, phenolics play a defensive role against pests and influence insect growth and feeding which has high levels of flavonoids and antibiosis mechanisms that prevented Spodoptera litura larval growth (Mallikarjuna et al. 2004). Found plants that contain a high level of phenolic compounds were less infested by the cereal aphid (Wójcicka 2010). Nitrogen (N), phosphorus (P), and potassium (K) are the major elements in optimizing potato yield. We examined the effect of our tested CEs and NEs on the NPK plant's content. Our data indicated all treatments decreased plant nitrogen contents. There was a significant increase in phosphorus content with plants treated with ECE and ENE followed by PCE and PNE. this means eucalyptus emulsions have a positive effect on phosphorus plant contacts. data of (Chrysargyris et al. 2020) agreed with this study. Where the application of essential oils of rosemary and eucalyptus affected the nutrient content in tomato leaves. Therefore, N leaf content decreased. While K, as well as P content, increased. Potato plants treated with PCE exhibited increases in potassium contents. The literature highlighted the importance of potassium evidence in obtaining high potato tuber yields (Kumar et al. 2007). our tested NEs showed a positive effect on plant potassium content. So, it may promote tubers' growth more than untreated plants. Through a greenhouse study, PNE did not show any negative effect on tomato plant health (Pandey et al. 2020). Foliar spray of soybean plants with thymol NE showed significantly higher values of plant height, root length, root fresh weight, number of nodules /plants, weight/nodule, number of pods /plant, and 100 seed weight as compared to control (water treatment), bulk thymol treatment. Peroxidases using H2O2 oxidize several substances. I considered peroxidases the plant defense-responsive system in plants. The biological effect of essential oils and their constituents causes oxidative by the accumulation of reactive oxygen species (Mousavizadeh and Sedaghathoor 2011). Based on the antioxidant property of essential oils, we evaluated the effect of both PNE and CNE on the increased peroxidase activity in potato leaves. Also, both NEs showed antioxidant activity higher than CEs in plant leaves. So, Peroxides act as defense enzymes against whitefly insects in potato fields. Our results agree with those obtained by (Ben-Jabeur et al. 2015) thyme oil applied on leaves of tomato plants induces plant systemic resistance by causing peroxidase accumulation. PCE and ECE increase plant Peroxidases so these oils may provide insect-resistant plants against B. tabaci on potato fields. We also evaluated NEs as promoting growth, as they can give significant effects on plant seedlings, either as promoting plant growth. We found the NE derived from peppermint EO could suppress fungi Alternaria solani growth while promoting tomato plant growth (Pandey et al. 2020). Thymol oil NE exhibited antibacterial activity and promoted soybean plant growth (Kumari et al. 2018).

Using novel pest management control methods for controlling B. tabaci, has become crucial to maintaining sustainable agricultural practices in Egypt essential oils course and nano-emulsions such as peppermint and eucalyptus oils offer a non-chemical, alternative solution to control these pests in the potato fields. While showing promising efficacy, we must monitor applications with the weather. These findings offer growers a practical and much-needed control option for maintaining potato crops, as widely used insecticide programs have decreased efficacy because of the increased application of conventional insecticides. But, the main disadvantage of using essential oils as an effective method for controlling insects is the unstandardized components of these essential oils. Therefore, additional research on plant cultivation and essential oil standardization is necessary.

Ethics approval and consent to participate

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Availability of data and materials

All data and materials are mentioned in the manuscript

Competing interests

The authors declare that they have no competing interests

Funding

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Authors’ contributions

All authors designing carried out the experiments,MNW and EAAreared B. tabaci,TFW collected the data analyzed the data,MNW and TFW wrote the manuscript. all authors read and approved the final manuscript

Acknowledgements

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Competing interests

The authors have no competing interests to declare that are relevant to the content of this article.

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Table (1): chemical component (%) of peppermint and eucalyptus essential oils

Components	% P	% E	RT (min)
3-Methyl-apopinene	2.04		5.87
alpha-Terpinene	-	19.78	6.37
Tetrahydro-m-cresol	0.52		6.49
Camphene	-	0.06	6.81
Sabinene	3.25		7.28
Menthen	0.43	0.96	7.80
Trans-Carane	0.73		7.95
áMyrcene	-	0.69	8.08
alpha-Phellandrene	-	0.63	8.53
Phenelzine	-	0.13	8.96
1H Pyrazole, 1 vinyl	-	1.12	9.23
1,8-Cineole	11.26	27.78	9.45
β-cis-Ocimene	-	0.07	9.98
çTerpinene	-	5.09	10.35
1,2-Oxolinalool	-	0.06	10.91
(+)2Carene	-	0.47	11.29
(+)-Linalool	-	0.21	11.79
Menthone	32.53	-	11.94
exo-Fenchol	-	0.15	12.33
1-Terpinenol	-	0.40	12.95
Isopinocarveol	-	0.40	13.16
Isoborneol	-	0.24	14.05
Terpinen4ol	-	0.64	14.29
1-p-Menthol	-	37.24	14.86
Camphene	-	2.34	15.02
Menthol	35.90	-	16.07
Pulegon	1.53	-	18.61
Longifolene	-	0.24	20.69
Cyclohexanol, 5-methyl-2-(1-methylethyl) acetate, (1à,2á,5á)-	7.53	-	21.12
Caryophyllene	4.29	0.06	26.40
nHexadecanoic acid	-	0.88	33.07
13-Octadecenoic acid	-	0.66	33.50
Z-(13,14-Epoxy) tetradec-11-en-1-ol acetate	-	0.83	34.14
Carane	-	0.23	34.29
Monoelaidin	-	13.39	34.38
Octadecanoic acid	-	0. 98	34.94
alpha-Linoleic acid	-	0.59	35.13
Benzyl oleate	-	0.16	35.26
Heptacosane	-	0.78	35.45
Octadecanedioic acid	-	0.22	35.77

Table (2): The characterization of the nanoemulsion Peppermint oil and Eucalyptus oil formulations

Treatments

particle size

(nm)

polydispersity index (PDI)

Viscosity

(cp)

PNE

114.5

0.27

0.88

ENE

145.8

0.69

0.88

Table (3): Efficacy of P (CE, NE) and E (CE, NE) against whitefly on potato field during season 2021.

Treatments	Mean No of nymph/plant before treatment (± SE) (1 Nov.)	Mean percent reduction of nymph (± SE)
		After the first spray			After the second spray
		3 days (4Nov.)	7 days (7 Nov.)	10 days (10 Nov.)	3 days (13 Nov.)		7 days (17 Nov.)		10 days (20 Nov.)
Control	15.00±2.88	7.96±2.00d	9.52±4.76c	8.06±3.57c	11.97±5.20d	10.21±2.56c		12.91±5.14d
PCE	17.33±2.66	83.13±1.43ab	71.13±1.52ab	67.12±0.38a	82.64±5.18ab	79.09±6.49a		69.71±1.94b
PNE	18.66±4.37	82.09±094b	68.42±4.55ab	60.65±7.15ab	84.14±0.24a	76.80±2.14a		68.95±0.48bc
ECE	16.00±6.00	71.09±0.46c	67.66±0.75ab	57.61±2.98ab	73.26±0.86bc	66.68±1.74b		67.00±0.62bc
ENE	15.66±3.48	72.60±0.00c	65.23±1.61b	54.96±1.64b	72.72±0.83c	63.60±1.52b		61.90±0.87c
Imidacloprid	16.33±1.33	88.31±2.66a	77.28±1.80a	67.24±0.56a	89.03±1.26a	83.22±1.67a		78.52±2.47a

*Means followed by the same letter are not significantly different (LSD test at ˂0.05).

Table (4): Efficacy of P (CE, NE) and E (CE, NE) against whitefly on potato field during season 2022.

*Means followed by the same letter are not significantly different (LSD test at ˂0.05).

Treatments	Mean No of nymph/plant before treatment (± SE)	Mean reduction% of nymph (± SE)
		After the first spray			After the second spray
		3 days (4Nov.)	7 days (7 Nov.)	10 days (10 Nov.)	3 days (13 Nov.)	7 days (17 Nov.)	10 days (20 Nov.)
Control	23.16±4.14	18.21±5.22c	8.99±5.08c	9.68±2.84d	5.84±1.73d	9.64±3.11c	13.69±4.79c
PCE	25.16±2.28	79.88±5.02ab	73.66±2.01ab	66.17±6.32ab	78.58±1.76b	74.29±2.11ab	65.76 ±3.70b
PNE	26.66±3.49	76.96±3.39ab	73.20±1.02ab	65.09 ±2.66ab	83.27±3.75ab	81.78±5.47a	71.81±2.34ab
ECE	25.83±2.79	73.36 ±0.62b	67.49±1.95ab	53.16±0.93c	77.85±2.01b	70.99±0.00b	66.06±3.39b
ENE	26.00±3.39	68.78±1.22b	65.30±4.21b	55.99±2.26bc	69.97±2.92c	66.16±3.07b	64.37±3.23b
Imidacloprid	25.83±2.79	87.22±1.94a	75.85±3.72a	73.21±3.13a	89.26±0.67a	82.27±3.84a	77.94±3.39a

Table (5): Effects of treatments and their interactions on whitefly nymphs in 2021 and 2022.

	2021				2022
	F value	df	P value	LSD _0.05	F value	df	P value	LSD _0.05
spray	4.970	1	0.0457*	3.007	21.724	1	0.0005***	1.877
treatment	124.56	5	0.0000***	7.231	213.811	5	0.0000***	5.610
time	44.177	2	0.0000***	2.291	62.737	2	0.0000***	2.162
time*spray	3.385	2	0.0421*	3.239	3.755	2	0.0305*	3.057
spray * treatment	1.835	5	0.1813ns	7.336	0.697	5	0.6358ns	7.726
time*treatment	2.113	10	0.0415*	2.145	3.281	10	0.0026**	5.296
treatmentspraytime	1.164	48	0.337ns	7.934	1.01	48	0.4449ns	7.488

Table (6): Effects of Peppermint and Eucalyptus oils course and nano emulations on growth components and Peroxidases of potato plant in the field

Treatments	Mean total proteins (mg/ g fresh weight) ± SE	Mean total carbohydrates (mg/ g fresh weight) ± SE	Mean total phenols (mg/ g fresh weight) ± SE	Mean nitrogen (mg / g fresh weight) ± SE	Mean phosphorus (mg /g fresh weight) ± SE	Mean potassium (mg /g fresh weight) ± SE	Peroxidases (∆ O.D. /min/ g fresh weight) Mean± SE
Control	11.13a±0.44	47.53b±1.31	1.92b±0.06	2.01a±0.24	0.53c±0.05	1.81c±0.11	474.33c±8.68
PCE	9.73b±0.14	49.56b±2.20	1.83b±0.03	1.96b±0.18	0.58bc±0.02	3.11a±0.19	521.1cb±13.01
PNE	10.73a±0.32	50.93ab±50.93	1.83b±0.02	1.79b±0.19	0.59bc±0.01	2.47b±0.12	570.33ab±9.83
ECE	9.73b±0.21	57.1a±2.15	1.88b±0.02	1.67b±0.16	0.71a±0.02	2.00c±0.05	515.33cb±13.69
ENE	10.96a±0.17	52.2ab±1.51	2.06a±0.04	1.81b±0.17	0.64ab±0.02	2.47b±0.07	621.00a±14.73

*Means within each Columns followed by the same letter are not significantly different (LSD test at ˂0.05).

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Efficiency of essential oils emulsions against Whitefly Bemisia tabaci (GENN.) infesting potato plants under field conditions In Egypt

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Abstract

Figures

Introduction

Methods

Tested materials

GC-MS analysis

Emulsion preparation

Experimental area

Sampling methods

Estimation of total soluble proteins

Estimation of total carbohydrate

Estimation of total phenolic contents

Estimation of peroxidase enzyme activity

Statistical Analyses

Results

Chemical composition of peppermint and eucalyptus essential oils

Nano-emulsions characterization

Weather data

Effects of tested emulsions peroxidase enzyme

Discussion

Conclusion

Declarations

References

Tables

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