Insecticidal activity of crude extracts of four plants and their Ag Nano liquids against Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

doi:10.21203/rs.3.rs-1305717/v1

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Insecticidal activity of crude extracts of four plants and their Ag Nano liquids against Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

https://doi.org/10.21203/rs.3.rs-1305717/v1

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A good quantity of global agricultural production is being devastated due to insect pests which need serious attention by the agricultural scientists and policy makers. The sole dependence on agrochemicals has increased over the past decades. However, adoption of botanical insecticides is gaining momentum as an ecofriendly alternative to the hazardous chemical pesticides. Applications of bionanoparticles are highly effective and may limit the risk of synthetic pesticide usage. Leaf dip and topical application toxicity bioassay were carried out to investigate the biological effects of aqueous leave extracts of Annona squamosa, Justicia adathoda, Ipomea carnea and Pongamia glabura at different concentrations (1.25, 2.5, 5 and 10%) against first, second, third nymphal instars and adult of Phenacoccus solenopsis (Hemiptera: Pseudococcidae). The result suggest that the aqueous leaf extract of P. glabura (LC₅₀ = 9.673%) showed more impact after 144 hrs of exposure followed by J. adathoda, I.carnea and A. squamosa in leaf dip toxicity bioassay. In the topical application, P. glabura (LC₅₀ = 1.042%) recorded the highest insecticidal activity followed by other three tested plants. The AgNP’s of P. glabura-AgNP’s showed highest impacts (LC₅₀ = 4.750%) than other three tested nanoparticles i.e., I.carnea-AgNP’s, J. adathoda-AgNP’s, A. squamosa-AgNP’s in addition to Vijay neem, respectively. From these experiments, it is evident that the P. glabura-AgNP’s and J.adathoda-AgNP’s possess more insecticidal activity and hence these products could be used for P. solenopsis management.

Crude extract

AgNP’s

vijaya neem

Cotton Mealybug

The cotton mealy bug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is a polyphagous insect pest. It feeds on more than 154 plant species including field crops, vegetables, ornamentals, fruit trees, weeds and bushes and causes economic damage mainly to vegetables, oilseed and ornamental plants.^1,2 The infestations with mealy bugs on different host plants could be effectively controlled using biological control agents.^1,3−5

The evaluation of plant extracts for their deleterious effects on insects is one of the approaches used for the search of novel botanical insecticides.⁶ Botanical pesticides are biological products which are used as mixtures for preventing, fighting and killing a variety of pests such as weed, insects, rodents and microbes.⁷ Botanical pesticides are biodegradable⁸ and their use in crop protection could be an effective and sustainable alternative to synthetic chemicals. Azadirachta indica, Ocimum sanctum, Calotropis gigantea, Nicotina tabacum and Alium sativum⁹, Thymus vulgaris L. (Lamiaceae) (Whole plant), Artemisia absinthium L. (Asteraceae) (Seeds), Pluchea dioscoridis L. (Asteraceae) (Leaves), Cyperus articulatus L. (Cyperaceae) (Tubers), Mentha longifolia (L.) Huds. (Lamiaceae) (Leaves), Anethum graveolens L. (Apiaceae) (Aereal parts), Lantana camara L.(Verbenaceae) (Leaves), Zingiber officinale Rosc. (Zingiberaceae) (Roots), Elettaria cardamomum Maton (Zingiberaceae) (Seed pods), Syzygium aromaticum (L.) Merr. & Perry (Myrtaceae) (Buds)¹, Mentha longifolia, Mentha piperita, Mentha spicata, and Nepeta cataria¹⁰, A. indica, Eucalyptus globules, Ocimum basilicum L.¹¹ were used for the management of P. solenopsis. Further, neem oil and NSKE ¹², biopesticides and biorationals insecticides ¹³ or commercial need product (Neemosal)¹⁴ were also tested against P. solenopsis.

Proposed plants like Annona squamosa¹⁵, Justicia adatoda¹⁶, Ipomea carnea^17,18 and Pongamia glabra (= Pongamia pinnata)^11,19 showed insecticidal, repellent and antifeedant properties against various insect pests have been known for three decades. However, crude extracts of these plants have not tested against various life stages of P. solenopsis.

The nanotechnology has become one of the most promising new approaches for insect pest control.²⁰. Biological synthesis of nanoparticles has received increased attention due to a growing need to develop environmentally benign technologies in material synthesis.²¹ Various plant-based silver nanoparticles have been utilized in pest control like Nerium oleander ²², Sargassum muticum²³, Avivennia marina²⁴, Ficus religiosa and Ficus benghalensis²⁵, Punica granatum²⁶, Euphorbia prostrate.²⁷. It was also stressed that silver nanoparticles (AgNPs) synthesized by using tested essential oils of Cymbopogon citratus, Thymus vulgaris and Pelargonium graveolens and were evaluated against female of P. solenopsis. ^{28, 29}

The literature review shows that so far no work has been undertaken on the utilization of A. squamosa, J. adatoda, I. carnea and P. glabra against any life stages of P. solenopsis either under laboratory or field conditions. Therefore, considering the lacuna, the present study attempts to evaluate the efficacy of some native botanicals against cotton mealy bug, P. solenopsis. The present study was aimed to develop a rapid and simple biosynthesis of silver nanoparticle by reducing the metal (AgNO₃), using the leaf extracts. Hence with the following objectives: 1) impact of crude extracts of Annona squamosa, Justicia adathoda, Ipomea carnea and Pongamia glabura) and their bionanomaterials and commercial pesticides vijay neem on the mortality of P. solenopsis life stages and 2) to determine the lethal concentration (LC₃₀, LC₅₀ and LC₉₀) for the crude extracts and their bionanomaterials of selected botanicals and the vijay neem on the mortality of P. solenopsis life stages.

Collection of botanicals. Permission has been obtained for collecting the botanicals namely Annona squamosa L., (Family: Annonaceae) was collected from Palayapettai, Tirunelveli (N 08° 73’ 70.75” latitude andE 077° 66’ 57.73” longitude) (Plate 1.1a), Justicia adathoda L., (Family: Acanthaceae) was collected from St. Xavier’s College campus, Palayamkottai, Tirunelveli (N 08° 71’ 80.23” latitude and E 077° 73’ 81.22” longitude) (Plate 1.1b), Ipomea carnea J.(Family: Connolvulaceae) was collected from the Tirunelveli Town (N 08° 73’ 05.34” latitude and E 077° 68’ 75.15” longitude) (Plate 1.1c) and Pongamia glabura Ventenat. (Family: Papilionaceae) was collected from St. Xavier’s College campus, Palayamkottai, Tirunelveli ( N 08° 71’ 74.48” latitude and E 077° 74’ 01.66” longitude) (Plate 1.1d) Tamil Nadu, India. and The collected plant materials were washed twice with fresh water and ones with distilled water to remove the dust and debris.

Preparation of botanical crude extracts. Five gram of fresh and healthy leaves was chopped into small pieces and was taken in a 250 ml conical flask. 100 ml of distilled water was added to it and was heated in a water bath at 72°C for 1 hour. The decoction was then cooled and filtered through Whatman No. 1 filter paper. The filtrate was collected in a 50 ml sterilized standard flask and was used for further studies. All experimental protocols involving plant materials were conducted in accordance with institutional, national, and international guidelines and legislation.

Preparation of silver nanoparticles. Silver nitrate was purchased from Hi-Media (RM409-25G) with 99.9% purity was used for the preparation. 17 mg silver nitrate was taken and dissolved in the 100 ml of double distilled water (10^− 3M AgNo₃). To induce the synthesis of silver nanoparticles, 20 µl of plant extract was added at time interval 1 minute regularly. The finally synthesized pale yellow to reddish brown color and its store for 100 ml in sterilized standard flask used for this experimental study.

Preparation of commercial Vijay neem. Indian farmers have been using a neem- based insecticides Vijay neem (Madras Fertilizers Limited, Chennai) was purchased from local agrochemical market and field recommended dose of 0.03% was prepared (30 µl /100 ml water ) and used for this experiment.

Collection and rearing of Phenacoccus solenopsis. Life stages of P. solenopsis were collected from cotton field of Palayapettai (N 08° 73’ 70.75” latitude and E 077° 66’ 57.73” longitude) and Savalaperi (N 08° 49’ 55.6” latitude and E 077° 51’ 34.8” longitude) Tirunelveli, Tamil Nadu, India. The collected P. solenopsis cultures were maintained on cotton plant and also using newly sprouted potatoes shoots in plastic tray (height 30 cm × 24 cm width, depth 5 cm) under the laboratory conditions ( height 185 cm × 89 cm width) at room temperature (29 ± 2º C), 70–80% RH and 11 L: 13 D hours.⁹ The laboratory emerged insects were also maintained continuously for the experiment. Laboratory emerged first, second, third instar nymphs and adult P. solenopsis were used for the experimental study.

Toxicity bioassay -Leaf dip method. Leaf dip method was performed in plastic containers (4 cm diameter and 5 cm height). Four different concentrations (1.25, 2.5, 5 and 10%) of the aqueous extracts of used for the leaf dip method of toxicity bioassay. Fresh and healthy cotton leaves (KC-II) was cut chopped into small pieces of 2×2 cm size and then soaked for five minutes in the experimental solutions and were then shade dried for ten minutes.^46,47 The shade dried leaves were fed to the nymphs and adults of P.solenopsis. The six replications each with ten nymphs, adult per treatment were used for the experiment. Control category was treated with distilled water and teepol (100 ml + 50 µl). The animals were maintained as mentioned above. Mortality was recorded after 24, 48, 72, 96, 120 and 144 hrs and corrected according to the following formula.

% of the animals killed in treated - % of the animal killed in the control

Corrected percent mortality (CPM) = --------------------------------------------------------------------------------------------- × 100

100 - % of the animals killed in the control

Toxicity bioassay -Topical application. All the four different concentrations of the experimental solutions were sprayed separately on different sets of P.solenopsis nymphs and adults using a hand sprayer (capacity of 100 ml). The experimental animals were then introduced into the experimental rearing tray (23 cm width; 30 cm length and 5 cm height). Observations were made on the mortality of the animals at 24 hours intervals up to 144 hours after treatment (24, 48, 72, 96, 120 and 144 hrs).⁴⁸

Statistical analysis. Corrected mortality of insect was calculated using Abbott correction formula for natural mortality in untreated control.⁴⁹ The corrected mortality data was subjected to further analysis using SPSS.⁵⁰. The probit analysis (LC₃₀, LC₅₀ and LC₉₀), df-values, chi-square values and significance values were recorded. The mortality data were subjected to one way Analysis of Variance (ANOVA); the mean values compared by Tukey test (P = 0.005 and ‘P’ values arrived at to assess the statistical significance of values less than 0.005 were considered as significant using statistical package.⁵⁰

Insecticidal activity of crude extracts. Results of the experiments shows that significantly highest nymphal mortality was observed (df = 4,10; F = 12.659; p < 0.005; LC₅₀ = 9.673% and df = 4,10; F = 22.861; p < 0.005; LC₅₀ = 1.042% for leaf dip bioassay and topical application toxicity bioassay respectively) in P. glabura treated insects at 144 hrs against first instar nymphs of P. solenopsis (Tables 1 and 2; Fig. 1). In the second nymphal instar, leaf dip toxicity bioassay showed the highest nymphal mortality in P. glabura (93.33%; df = 4,10; F = 95.583; p < 0.005 an LC₅₀ = 5.145%) on 144 hrs against second nymphs instar P. solenopsis. However, topical application toxicity bioassay show that the highest nymphal mortality was recorded in J. adathoda on 144 hrs (76.66%; df = 4,10; F = 12.855; p < 0.005 and LC₅₀ = 3.295%) (Table 2, Fig. 2).

Table 1

Probit analysis of leaf dip bioassay using plant extracts on *P.solenopsis.*
Plant Extract	Life stages	LC₅₀	Chi-square	Significance
A.squamosa	First instar	1.245	1.797	0.407
	Second instar	1.495	0.894	0.344
	Third instar	8.277	3.561	0.059
	Adult	2.318	0.005	0.996
J.adathoda	First instar	7.408	1.326	0.515
	Second instar	1.376	0.575	0.448
	Third instar	1.366	3.255	0.071
	Adult	1.716	1.062	0.303
I.carnea	First instar	8.036	1.277	0.528
	Second instar	1.429	0.822	0.365
	Third instar	1.488	1.223	0.269
	Adult	1.892	0.441	0.506
P.glabura	First instar	9.673	0.06	0.970
	Second instar	5.145	0.874	0.350
	Third instar	4.541	0.316	0.574
	Adult	1.193	0.371	0.542

Table 2

Probit analysis of topical application bioassay using plant extract on *P.solenopsis*
Plant Extract	Life stages	LC₅₀	Chi-s quare	Significance
A.squamosa	First instar	1.139	1.567	0.457
	Second nstar	5.896	1.014	0.602
	Third instar	4.248	1.36	0.506
	Adult	6.317	2.858	0.204
J.adathoda	First instar	3.534	0.829	0.661
	Second nstar	3.295	0.656	0.72
	Third instar	4.201	1.298	0.523
	Adult	5.137	0.244	0.885
I.carnea	First instar	1.242	1.797	0.407
	Second nstar	7.408	1.326	0.515
	Third instar	8.036	1.277	0.528
	Adult	9.673	0.06	0.97
P.glabura	First instar	1.042	0.879	0.644
	Second nstar	8.925	0.506	0.777
	Third instar	5.863	1.435	0.488
	Adult	8.330	0.678	0.713

In the leaf dip toxicity bioassay of the laboratory experiment shows that the highest nymphal mortality was observed in P. glabura (90.00%; df = 4,10; F = 38.643; p < 0.005 and LC₅₀ = 4.541%) on 144 hrs followed by J. adathoda (76.66%; df = 4,10; F = 13.617; p < 0.005 and LC₅₀ = 1.366%), A. squamosa (76.66%; df = 4,10; F = 10.375; p < 0.001 and LC₅₀ = 8.277%) (Fig. 3) and I.carnea (66.66%; df = 4,10; F = 20.088; p < 0.005 and LC₅₀ = 1.488%) (Fig. 4) against third instar nymphs of P. solenopsis. But in the topical application bioassay the highest nymphal mortality was recorded in J. adathoda (df = 4,10; F = 15.967; p < 0.005 and LC₅₀ = 4.201%) followed by P. glabura (df = 4,10; F = 24.328; p < 0.002 and LC₅₀ = 5.863%) against third instar nymphs of P. solenopsis, after 144 hrs of exposure.

In leaf dip toxicity bioassay the highest adults mortality was recorded in P. glabura (100%; df = 4,10; F = 85.812; p < 0.005 and LC₅₀ = 1.193%) on 144 hrs against adult of P. solenopsis. The result of topical application toxicity bioassay presented in the Table 2 and Fig. 2 recorded the highest nymphal mortality in J. adathoda (83.00%; df = 4,10; F = 53.056; p < 0.005 and LC₅₀ = 5.137%) on 144 hrs.

The overall comparison of the nymphicidal activity of the experimental crude extracts using the cumulative average mortality of first, second, third nymphal instars and adult P. solenopsis the leaf dip method (86.666%) recorded the highest mortality in P. glabura followed by J. adathoda (78.33%), A. squamosa (71.66%) and I. carnea (69.99%) and the topical application bioassay recorded the maximum mortality (77.49%) in J. adathoda followed by P. glabura (73.33%), A. squamosa (62.49%) and (56.66%) in I.carnea (Table 3)

Table 3

Cumulative average mortality (%) of first, second, third nymphal instar and adult *P.solenopsis*.
Botanicals/AgNP’s	Average (%)
Botanicals/AgNP’s	Leaf dip method	Topical application method
A.squamosa	71.66	62.49
J.adathoda	78.33	77.49
I.carnea	69.99	56.66
P.glabura	86.66	73.33
A.squamosa – AgNP’s	86.66	88.33
J.adathoda– AgNP’s	90.83	89.99
I.carnea– AgNP’s	85.83	79.66
P.glabura– AgNP’s	95.83	86.66
Vijay neem	86.24	76.66

Insecticidal activity of AgNP’s. Insecticidal activity of AgNP’s on the first nymphal instar of P. solenopsis mortality was recorded for the experimental nanoparticles using in leaf dip and topical application toxicity bioassays (Tables 3 and 4; Figs. 5–8). Result of the experiments presented in Table 3, 4 and Figs. 5–8 shows that the highest nymphal mortality was observed in P. glabura- AgNP’s (86.66%; df = 4,10; F = 28.278; p < 0.005 and LC₅₀ = 4.750%) (Fig. 5) on 144 hrs in leaf dip method of toxicity bioassay against the first instar nymphs of P. solenopsis. However, in topical application toxicity bioassay showed highest nymphal mortality (90.00%; df = 4,10; F = 87.357; p < 0.005 and LC₅₀ = 1.276%) in A. squamosa- AgNP’son 144 hrs (Fig. 6).

Table 4

Probit analysis of leaf dip bioassay using bionanoparticles on *P.solenopsis.*
Bionano particles	Life stages	LC₅₀	Chi-square	Significance
AS-AgNps	First instar	1.734	0.122	0.941
	Second instar	6.433	0.36	0.849
	Third instar	1.491	0.792	0.373
	Adult	1.892	0.441	0.506
JA-AgNps	First instar	1.151	1.637	0.441
	Second instar	6.440	0.318	0.573
	Third instar	1.188	0.105	0.764
	Adult	1.641	2.147	0.143
IC-AgNps	First instar	5.951	0.033	0.984
	Second instar	9.029	0.373	0.541
	Third instar	1.714	0.635	0.425
	Adult	3.577	8.276	0.005
PG-AgNps	First instar	4.750	0.141	0.932
	Second instar	4.928	8.077	0.004
	Third instar	1.434	0.837	0.36
	Adult	1.894	0.681	0.409
Vijay neem	First instar	4.949	1.394	0.498
	Second instar	9.601	0.831	0.362
	Third instar	1.466	0.160	0.690
	Adult	6.812	0.273	0.601

In the second instars, significantly highest nymphal mortality (100%; df = 4,10; F = 86.75; p < 0.005 and LC₅₀ = 4.928%) in P. glabura-AgNP’s in leaf dip toxicity. Similar results was also recorded in the topical application toxicity test (df = 4,10; F = 219.167; p < 0.005 and LC₅₀ = 1.141%). Similar results was also observed for the third instar nymphs of P. solenopsis (df = 4,10; F = 52.293; p < 0.005 and LC₅₀ = 1.434%) when P. glabura-AgNP’s applied as leaf dip bioassay (Table 4). But the topical application bioassay showed the highest nymphal mortality (df = 4,10; F = 124.7; p < 0.005 and LC₅₀ = 1.434%) in J. adathoda-AgNP’s (Table 5; Fig. 7).

Table 5

Probit analysis of topical application bioassay using bionanoparticles on *P.solenopsis*.
Bionano particles	Life stages	LC₅₀	Chi-square	Significance
AS-gNps	First instar	1.276	4.871	0.088
	Second instar	1.226	2.258	0.323
	Third instar	1.508	2.17	0.338
	Adult	1.848	0.094	0.954
JA-AgNps	First instar	1.034	0.106	0.948
	Second instar	1.141	0.996	0.608
	Third instar	1.434	2.667	0.264
	Adult	1.795	0.414	0.813
IC-AgNps	First instar	1.734	0.122	0.941
	Second instar	1.151	1.637	0.441
	Third instar	5.951	0.033	0.984
	Adult	4.750	0.141	0.932
PG- AgNps	First instar	7.399	1.328	0.515
	Second instar	4.229	0.013	0.993
	Third instar	8.812	0.028	0.986
	Adult	3.738	1.133	0.568
Vijay neem	First instar	7.048	3.19	0.203
	Second instar	1.076	7.522	0.023
	Third instar	4.949	1.394	0.498
	Adult	6.595	1.886	0.397

Result of the leaf dip toxicity bioassay recorded significantly highest (df = 4,10; F = 0.886; p < 0.005 and LC₅₀ = 1.894%) adult mortality in P. glabura -AgNP’s. Similar results was also recorded for topical application toxicity bioassay (df = 4,10; F = 76.062; p < 0.005 and LC₅₀ = 3.738%) for P. glabura -AgNP’s. I.carnea -AgNP’s (86.66%; df = 4,10; F = 28.278; p < 0.005 and LC₅₀ = 4.750%) showed least impacts to the adult of P.solenopsis (Fig. 8).

Overall comparison of nymphicidal activity of the experimental AgNP’s using the cumulative average mortality of first, second, third nymphal instar and adult P. solenopsis the leaf dip toxicity bioassay using P. glabura-AgNP’s (95.83%) followed by J. adathoda-AgNP’s (90.83%), A. squamosa-AgNP’s (86.66%) and I. carnea-AgNP’s (85.83%) and followed by topical application bioassay recorded in J. adathoda-AgNP’s (89.99%), A. squamosa-AgNP’s (88.33%), P. glabura-AgNP’s (86.66%) and (79.66%) in I. carnea-AgNP’s (Table 3).

Insecticidal activity of vijayneem. The mortality was recorded for experimental sample vijayneem using in leaf dip and topical application toxicity bioassays. Results of the experiments presented in the (Fig. 9) shows that the highest nymphal mortality (80.00%; df = 4,10; F = 39.5; p < 0.005 and LC₅₀value = 4.949%) was recorded in vijayneem on 144 hrs in leaf dip toxicity bioassay followed by first instar nymphs of P. solenopsis and (83.33%; df = 4,10; F = 34.726; p < 0.005 and LC₅₀ = 9.601%), against second instar nymphs of P. solenopsis and followed by (81.66%; df = 4,10; F = 24.236; p < 0.005 and LC₅₀ = 1.466%), against third instar nymphs of P. solenopsis and then (100.00%; df = 4,10; F = 61.837; p < 0.005 and LC₅₀ = 6.812%), against adult of P. solenopsis.

Figure 9 shows that the highest nymphal mortality (80.00%; df = 4,10; F = 41.167; p < 0.005 and LC₅₀ = 7.048%) was observed in vijayneem on 144 hrs in topical toxicity bioassay followed by first instar nymphs of P. solenopsis and then (50.00%; df = 4,10; F = 19.206; p < 0.001 and LC₅₀ = 1.076%), against second instar nymphs of P. solenopsis and followed by (80.00%; df = 4,10; F = 39.5; p < 0.005 and LC₅₀ = 4.949%), against third instar nymphs of P. solenopsis and adult (76.66%; df = 4,10; F = 31.179; p < 0.005 and LC₅₀ = 6.595%) of P. solenopsis.

Among the cumulative average mortality of vijayneem in leaf dip toxicity bioassay recorded (86.24%) mortality followed by topical application toxicity bioassay (76.66%) (Table 3). Over all the leaf dip and topical application toxicity bioassay of highest insecticidal activity was recorded in nanomaterials, vijayneem and followed by crude extract in high concentration (10%) by insecticidal activity of first, second, third nymphal instars and adult P. solenopsis.

The plants have evolved a plethora of secondary metabolites which play a major role in insecticidal activity.^16,30 The recent research was highly drifted on screening and characterization of green chemistry with potential target insecticidal agents.³¹ The recent studies have demonstrated the insecticidal properties of chemicals derived from plants that are active against specific target species, biodegradable to non-toxic products and potentially suitable for use in integrated pest management programs.³²

The AgNPs of PG, JA, AS and IC showed 100% mortality at various exposure periods, which indicates that there are lots of scope for reducing the nanoparticles level in P. solenopsis management and that this can be extended to other crops too. Results also shows does dependent mortality as observed for plant extracts to P. solenopsis. ^{1,33, 34} The present study also reveals that AgNPs synthesized using aqueous leaf extract of A. squamosa, J. adathoda, I. carnea and P. glabura showed insecticidal effects on P. solenopsis. The recent research on insecticidal action of plant materials specially secondary metabolites and essential oils resulted that they are eco-friently, biodegradable and species specific.^35–38

Compared with the LC₅₀ values was shown to be 3.4, 2.9, 2.2 and 2.0 times lower for AG-AgNPs, JA-AgNPs, PG-AgNPs and AC-AgNPs than for the A. squamosa, J. adathoda, P. glabura and I. carnea crude extract exposure. An opposite trend was recorded when essential oils of C. citratus, T. vulgaris and P. graveolen sAgNPs were evaluated against P. solenopsis.²⁹ Therefore, JA-AgNPs, AG-AgNPs, PG-AgNPs and AC-AgNPs were 3.7, 3.6, 1.8 and 1.4 times more toxic than the commercial formulation (vijaynem) respectively, which suggested that the biological activity was considerably improved through using AgNPs, and bionano materials have significantly better insecticidal toxicity than that of the crude extracts and commercial formulation. Moreover, bio-nanomaterials can save the manpower through decreasing the number of pesticide applications needed, and pesticide dosage compared to traditional pesticide. Furthermore, nanoparticles size of A. squamosa-AgNPs, J. adathoda-AgNPs, I. carnea-AgNPs and P. glabura-AgNPs is 1.56–3.13 nm, 1.56–3.13 nm, 1.56–3.13 nm and 1.56–3.13 nm respectively in AFM analysis. These small size can increase the penetration and absorption of the active ingredients by the pest.³⁸⁹⁻⁴¹

Many factors influencing the effectiveness of plant extracts and or bio-nano materials to control insect pests are different plant species, part of plant, tested insects, methods of extraction, type of solvents, and method of application. Plant extracts coupled with nanotechnology that are active at reducing pest populations at the insect stage can provide many associated benefits to pest control. AS, JA, IC and PG-AgNPs was considered to be potential agents for various biological applications including insecticidal agents.⁴² The results indicate that different concentration of all nanoparticles caused higher mortality in leaf dip method toxicity and topical application method toxicity bioassays. This indicates that the all-AgNPs in the intracellular space can bind to sulphur containing proteins or to phosphorus containing protein compounds like DNA, leading to the denaturation of some organelles and enzymes⁴³ causing death. Further, the mechanism which causes the death of the insect could be the ability of the nanoparticles to penetrate through the insect membranes^21,44 reported that half-life values for rotenone concentration with ZnO nanoparticle under UV radiation were found to be higher than those of rotenone alone. It may also be applied here, but it requires future studies. Under natural conditions, the released insect moved to various places for hiding during afternoons.

Insecticide based on Neem oil works as a systemic insecticide for many plants when applied as a soil drench. Once the product is in the plant’s vascular system, insects take it in during feeding. The commercial neem-based biopesticide at different concentration showed drastic mortality when combined with monocrotophos in contact toxicity. This proves the statement, “The chemical pesticides will continue to be the main weapon against P. solenopsis for the near future”. Mortality was varied according to the developmental stage, and type of bioassays used⁴⁵ as observed in the present study.

The present study illustrates that the botanicals A.squamosa, J.adathoda, I.carnea and P.glabura is a potential one for the eco-congenial nymphicide development as an alternate to chemical insecticides that are being currently used in cotton pest management programs. Our results reveal that the nymphal mortality and adult mortality is high in leaf dip method of toxicity bioassay followed by topical application. Mortality was enhanced while crude extracts combined with the bionanoparticles, vijay neem and they are considered most suitable for environment friendly pest management.Perhaps to reduce the quantity of chemical pesticides, farmers can be encouraged to use the concentrations for the pest management programme.

Acknowledgement

This research publication was financed by Taif University Researchers Supporting Project number (TURSP -2020/92), Taif university, Taif, Saudi Arabia.

Author contributions

Kitherian Sahayaraj: Conceptualization; Investigation; Methodology; Funding acquisition; Supervision; Project administration; Data curation; Writing - original draft; Writing - review & editing.

Madasamy, M : Investigation; Methodology; Data curation; Software, Visualization, Writing - original draft, review & editing.

Samy Sayed: Funding acquisition, Resources; Data processing; Writing - review & editing.

Hosny Kesba: Funding acquisition, Resources; Data processing; Writing - review & editing

Prathas Selvaraj: Conceptualization; Methodology; Co-Supervision; Project administration; Data curation; Writing - review & editing.

Madasamy, K: Data curation; Software; Visualization; Writing - review & editing.

Conflict interest statement

The authors declared they have no conflict of interest.

Data availability statement

The datasets generated and/or analyzed during the current study are not publicly available due reserved for filing a patent but are available from the corresponding author on reasonable request.

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Insecticidal activity of crude extracts of four plants and their Ag Nano liquids against Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

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