1. Introduction
The fall armyworm Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) causes serious damage to many plants of the Poaceae, Asteraceae, and Fabaceae families [1]. It is a serious agricultural pest that is native to tropical and subtropical areas of America. It has a wide host suitability range, disperses rapidly, and has currently invaded almost 100 countries around the world by quickly establishing itself in new ecosystems [2]. In Mexico, it is reported extensively, mainly in corn (Zea mays Linnaeus Poaceae) [3].
To counteract the damage caused to corn by S. frugiperda, its populations have been subjected to high pressure by the use of insecticides such as organophosphates, carbamates, and pyrethroids [4,5]. In the last 20 years, the use of synthetic chemical insecticides has predominated in control of S. frugiperda, especially of the N-oxide derivative benzofuroxan methyl-5-carboxylate N-oxide [6]. Unfortunately, excessive use of pesticides causes the loss and destruction of biodiversity [7]. It also leads to the increase in detoxification by microsomal oxidases, leading to the development of fall armyworm populations that are resistant to insecticidal compounds [8,9,10] such as chlorpyriphos (organophosphate), methomyl (carbamate), and lambda-cyhalothrin (pyrethroid) [11,12]. Given these problems, it is necessary to use other management alternatives against populations of S. frugiperda, for example, natural products isolated from plants with larvicidal activity [13,14,15].
The plants of the Ficus genus of the Moraceae family have developed various defense strategies against insect attack. These include physical defenses such as leaves with latex, hard leaves, mineralized leaves, and leaves with glandular trichomes [16,17,18], as well as several different metabolites and proteins that act against herbivores [19]. One recent study indicates that alkaloids from nine species of Ficus affect herbivorous insects [20]. Other research mentions that extracts from branches and leaves of Ficus spp. enriched in furanocoumarins serve as a defense against the neotropical brown stink bug, Euschistus heros (Fabricius) (Heteroptera: Pentatomidae). [21]. Our working group recently published the insecticidal potential of the ethanolic extract of Ficus petiolaris (Kunth) (Moraceae) against the aphid Melanaphis sacchari Zehntner (Hemiptera: Aphididae), and its pure compound 8-Methoxypsoralen, known as xanthotoxin, continued to present a toxic effect on this aphid [22]. In this context, the objective of this study was to evaluate the larvicidal effect of the ethanolic extract, its fractions, and the furanocoumarin 8-Methoxypsoralen of F. petiolaris on S. frugiperda larvae.
2. Materials and Methods
2.1. Plant Material
The stems and bark (6.115 kg) of F. petiolaris were collected in November 2022 from neighboring mature trees in the Sierra de Huautla Morelos Biosphere Reserve (REBIOSH) Mexico (Latitude: 18°23′49″ N, longitude: 96°12′30″ W. Altitude: 1083 m above sea level). For identification, a sample of F. petiolaris was deposited in the HUMO Herbarium of the Biodiversity and Conservation Research Center (CIByC) of the Autonomous University of the State of Morelos (UAEM). The identification was made by MSc. Gabriel Flores Franco with herbarium number HUMO-29538. The material was dried in the shade at room temperature (24 ± 2 °C) and ground with a mill (A 10 basic, IKA Works, Wilmington, NC, USA) to obtain particles of 5 to 10 mm (3.58 kg).
2.2. Extraction of Plant Material
The dried and crushed material of the stem and bark (2.5 kg) of F. petiolaris was extracted with ethanol (100%, J.T. Baker®, Thermo Fisher Scientific, Madrid, Spain). The process was carried out for three days and repeated three times using 5 L of solvent per kg of plant material. The plant residue was filtered using Whatman No. 5 paper (Merck KGaA®, Darmstadt, Germany). Subsequently, the ethanol was completely removed from the solution obtained by distillation under reduced pressure using a Büchi R-205 rotary evaporator (Equipar®, Ciudad de México, Mexico) then brought to complete dryness using a Model CEG-60 extraction hood (Industrias Figursa®, Estado de México, Mexico), to obtain the ethanolic extract of Ficus petiolaris (=EEFP, 82.9 g, yield of 3.3%) [22].
2.3. Purification of 8-Methoxypsoralen
Fifty grams of the ethanol extract was weighed and adsorbed 1:1 in silica gel (60 70-230, Merck KGaA®, Darmstadt, Germany) and suspended in n-hexane in a 95 × 4.2 cm length-diameter chromatographic column. The column was packed with 250 g of silica. The elution of the column began with 100% n-hexane, making polarity changes by adding acetone until ending with 100% acetone. We collected 231 fractions of 250 mL each. The fractions were combined using analysis by thin layer chromatography (TLC) with silica gel aluminum chromatographs (Kiessegel 60 F254, 20 × 20 cm × 0.2 mm, Merck), resulting in five sub-fractions of lower complexity than the original extract (group 1 of Ficus petiolaris-FpR1; group 2 of Ficus petiolaris-FpR2; group 3 of Ficus petiolaris-FpR3; group 4 of Ficus petiolaris-FpR4; group 5 of Ficus petiolaris-FpR5). The TLC analysis showed the presence of furanocoumarin in sub-fraction 2. Sub-fraction 2 (2.65 g) was absorbed with 7 g of SiO2 60 (70-230, Merck) diluted in n-hexane. Elution was performed with 100% n-hexane. Seventy-eight fractions of 15 mL each were collected. By TLC, 8 sub-fractions were obtained and with the help of an ultraviolet light lamp (Estela brand) from fractions 5–7 developed with 2% ceric ammonium sulfate, 8-Methoxypsoralen could be observed, from which it was possible to isolate 1461 mg. Structural elucidation was carried out using infrared (IR), hydrogen nuclear magnetic resonance (1H NMR), carbon thirteen (13C NMR) and mass spectrometry techniques (following [22] Figure 1).
2.4. Collection and Breeding of Spodoptera frugiperda
Spodoptera frugiperda larvae were collected from corn (Z. mays) crops in Yautepec, Morelos during July 2022. Diseased and parasitized larvae were eliminated in the CeProbi-IPN Entomology laboratory. Healthy larvae were analyzed for taxonomic characteristics to confirm the species S. frugiperda [23]. Healthy larvae were fed individually with artificial diet [24] contained in plastic vials. The insects were kept in a climate chamber at 25 ± 2 °C, 60 ± 5% RH and photoperiod 12:12 h L:D. The pupae were placed in waxed paper bags with a 10 cm diameter plastic Petri dish to emerge as adults. These adults were placed in waxed paper bags with a container of cotton moistened with a water + honey solution for feeding. These adults were mated randomly and laid masses of eggs that were collected and placed in Petri dishes with moistened cotton swabs. When the larvae hatched, they were given an artificial diet. The F2 larvae were used in the bioassays.
The lethal concentrations of extract (EEFP), fractions, and pure 8-Methoxypsoralen (xanthotoxin) causing 50% and 90% mortality in the population (LC50 and LC90, respectively) were determined through a diet ingestion bioassay. For the bioassays, the concentrations of each treatment were as follows: (1) Ficus petiolaris extract were 0, 500, 1000, 1500, 2000, and 2500 mg/kg of artificial diet (0, 500, 1000, 1500, 2000, and 2500 ppm), (2) fractions from Ficus petiolaris extract were 0, 250, 500, 750, 1000, and 1250 mg/kg of artificial diet (0, 250, 500, 750, 1000, and 1250 ppm), and (3) pure 8-Methoxypsoralen (xanthotoxin) were 0, 12.5, 25, 50, 75, 100 mg/kg of artificial diet (0, 12.5, 25, 50, 75, and 100 ppm). These concentrations used were determined from previous studies [25,26] with activity on larvae of this pest insect and other Noctuidae.
Five milliliters of treated diet were added to plastic containers measuring 3.0 × 3.5 cm in height and diameter and were allowed to solidify for 24 h. One F2 larva was placed in each plastic cup (feeding chamber), which was placed in a rearing chamber (Precision Brand, incubator 818, TEquipment®, Long Branch, NJ, USA) at 27 °C ± 1.5 °C, 60 ± 7% relative humidity, and a 12:12 h light/dark photoperiod. The treatments were evaluated in a simple randomized experimental design. A total of 30 neonate larvae were used per treatment where each larva was the experimental unit with each treatment having three replications, for a total of 90 larvae per treatment. The dependent variables were the larval weight decrease (7 days) and pupal weight decrease (total sclerotization) (mg), the proportion of larvae that pupated (% larval mortality), the proportion of pupae that did not emerge as moths (% pupal mortality), and the proportion of larvae + pupae (% cumulative mortality).
2.5. Statistical Analysis
A test of normality (Shapiro–Wilk W) and homoscedasticity (Bartlett test) was performed for all measured variables. One-way analysis of variance (ANOVA) and Tukey test (p < 0.05) were performed to identify potential differences between treatments using Statistix v. 8 (Analytical Software®, Tallahassee, FL, USA) [27]. Probit analysis was used to calculate the 50 and 90% lethal concentrations (LC50 and LC90) for larval and cumulative mortality, using the statistical analysis program BioStat 2.0 [28].
3. Results
3.1. Insectistatic Effect of Ethanol Extract of Ficus petiolaris
The analysis revealed a statistically significant difference between the treatment means (F(2, 6) = 646.67, p ≤ 0.001). The Tukey test found statistically significant differences between the artificial diet and the EEFP extract at 1500, 2000, and 2500 ppm (p ≤ 0.001), but no significant differences existed between the artificial diet and EEFP at 500 and 1000 ppm (p = 0.072). The weight of the larvae fed EEFP at 2500, 2000, and 1500 ppm was 89.5%, 89.1%, and 43.8% lower, respectively, than the weight of larva only fed with the artificial diet (diet = 40.14 mg, Figure 2A, Table S1). The same was true for pupal weight, with statistically significant differences between treatments (F(2, 6) = 51.72, p ≤ 0.001). The Tukey test found statistically significant differences between the artificial diet and EEFP extract at 2500, 2000, and 1500 ppm (p ≤ 0.001). These high EEFP concentrations reduced pupal weight by 17.9–20.0% (diet = 146.05 mg, Figure 2B).
3.2. Insecticidal Effect of Ethanol Extract of Ficus petiolaris
Table 1 shows the results of the cumulative mortality of the EEFP extract. There was a clear insecticidal effect of the extract on the larvae. The highest larval mortality occurred at a concentration of 2500 ppm (60%) (F (2, 6) = 616.74, p < 0.001), followed by concentrations of 2000 and 1500 ppm with mortality rates of 56 and 53%, respectively. Although there were statistically significant differences between the extract concentrations evaluated, they did not have an insecticidal effect on the pupae because they did not cause mortality ≥50%. When considering the cumulative mortality rate, the highest mortality was 80% with 2500 ppm, followed by the concentrations of 2000 and 1500 ppm, with mortality rates of 64% and 60%, respectively. No treatment was statistically similar to the positive control (Methyl Parathion®), which caused 100% larval mortality in just 24 h of exposure, while the negative control (artificial diet) had only 7% cumulative mortality (F(2, 6) = 287.96, p < 0.001, Table 1, Figure 3, Table S2). The EEFP extract had lethal concentrations of LC50 = 1396.9 ppm and LC90 = 3744.4.
3.3. Insectistatic Effect of the Fractions
Among all of the fractions evaluated, FpR4 presented the strongest insectistatic effect, decreasing the weight of the larva by 35.14–16.96% (from highest to lowest concentration evaluated; Table 2, Table S3). This was statistically significant different compared to the artificial diet control (diet: 43.73 mg, F(2, 26) = 68.02, p = 0.001). Similarly, the FpR5 fraction caused a reduction in larval weight by 23.37–10.70% (Table 2). With the FpR2 fraction at 1250, 1000, and 750 ppm, the weight of the fall armyworm larva was inhibited by 30.59, 23.32, and 15.82%, respectively (Table 2). Pupal weight decreased with the FpR4 fraction by 18.14–9.66% (F(2, 26) = 58.89, p = 0.001, Table 2), while the higher concentrations (1250, 1000, and 750 ppm) of the FpR5 fraction decreased the weight of the pupa by 9.0–6.83% (Table 2).
3.4. Insecticidal Effects of the Fractions
Among all of the fractions evaluated, FpR2 presented the strongest insecticidal effect, with a cumulative mortality of 100% and 93% at 1250 and 1000 ppm and an LC50 = 668.46 ppm and LC90 = 1138.46 ppm. This was statistically indistinguishable from the positive control methyl parathion (F(2, 26) = 871.24, p ≤ 0.001). The FpR1 fraction presented the second strongest insecticidal effect, with a cumulative mortality rate of 90% and 83% at 1250 and 1000 ppm and an LC50 = 857.8 ppm and LC90 = 2057.9 ppm. The positive control achieved 100% cumulative mortality in less than 24 h, and the negative control only eliminated 5% of S. frugiperda during the entire experiment (Table 2, Table S4).
3.5. Insectistatic Effect of 8-Methoxypsoralen
This furanocoumarin, which was isolated from the FpR2 fraction, presented the strongest insectistatic effect. All concentrations of 8-Methoxypsoralen decreased the weight of the larvae, which was statistically significant different from the negative control (diet = 47.53 mg; F(2, 6) = 239.96; p ≤ 0.001; Table 3, Table S5). The highest concentrations tested (100–75 ppm) inhibited the weight of the fall armyworm larva by 67.72 and 65.97% (Table 3). Furthermore, all concentrations of 8-Methoxypsoralen decreased pupal weight; the highest concentration tested (100 ppm) inhibited pupal weight by 24.08%, which was statistically significant different from the pupal weight of the negative control (diet = 148.25 mg; F(2, 6) = 310.22; p = 0.001; Table 3).
3.6. Insecticidal Effect of 8-Methoxypsoralen
The concentration of 100 ppm presented 100% larval mortality (LC50 = 67.68 ppm and LC90 = 505.17 ppm), which was statistically indistinguishable from the positive control, methyl parathion (F(2, 6) = 601.76; p ≤ 0.001; Table 3). The other concentrations also achieved important larvicidal effects, eliminating 64–96% of the fall armyworm larvae (Figure 4). The positive control eliminated 100% of S. frugiperda in just 24 h and the negative control only eliminated 8% throughout the experiment (Table 3, Figure 4).
4. Discussion
The higher concentrations of EEFP caused a decrease in body weight and elongation of developmental time in the larval and pupal stages of the pest insect. Antifeedant and antibiotic events may have caused these observed effects. Antifeedant or feeding deterrent behavioral effects causing larval growth inhibition have previously been reported for other noctuid species of economic importance [29]. Plants in diverse families can have secondary metabolites affecting several insect pests. These plants’ extracts can exhibit ovicidal, antifeedant, antigonadal, oviposition-deterrent, and repellent properties against different insect species [30], e.g., the extracts of the castor bean Ricinus communis (Linnaeus Euphorbiaceae) against S. frugiperda exhibited insectistatic activity [25].
In addition, EEFP was toxic by causing an insecticidal effect on the larvae, which increased with increasing concentration. This activity increased when considering its fractions and even more so with linear 8-Methoxypsoralen (xanthotoxin), which caused a greater inhibition effect on the larva, as well as the greater insecticidal effect on the fall armyworm at 100 ppm. Furanocoumarins have shown good insectistatic and insecticidal effects. The compound 8-Methoxypsoralen in a leaf disc test exhibited antifeedant activity on cotton leafworm Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) at concentrations of 500 to 1000 ppm [31]. Another study tested the phagodepressant effect of five furanocoumarins—bergapten, xanthotoxin, psoralen, imperatorin, and angelicin—against larvae of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae), finding synergistic antifeedant effects, i.e., when imperatorin and xanthotoxin, or bergapten and psoralen were used together, they had a stronger effect than the sum of the effects of each individual compound [32]. A study demonstrated that xanthotoxin (DC50 = 0.9 μg/cm2) had effective feeding deterrent and growth inhibitory effects in larvae (L3) of the false cabbage meter Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) [33]. Increasing concentrations of linear furanocoumarins prolonged larval development of T. ni. Furthermore, increasing concentrations of linear furanocoumarins increased T. ni mortality [34]. One study analyzed the toxicity of the linear furanocoumarin xanthotoxin (8-Methoxypsoralen) to the beet armyworm Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) under short ultraviolet (UVB) radiation; increasing xanthotoxin concentrations statistically significant decreased larval weight, extended generation time, and induced higher mortality [35]. When fed lineal foranocoumarin xanthotoxin throughout larval development, the survival of S. exigua colonies were sensitive to xanthotoxin exposure. In addition, the pupal weight and the egg production of those individuals surviving exposure did not indicate furanocoumarin resistance status interaction [36]. Furthermore, without causing effects of furanocoumarins on the larval-pupal parasitoid, Archytas marmoratus (Townsend) (Diptera: Tachinidae) of S. exigua in the presence and absence of ultraviolet radiation, there was no effect of increasing linear furanocoumarin levels on surviving parasitoid development time (from the time of host pupation) or size [37]
In another study, the insecticidal effect of xanthotoxin was demonstrated, and this effect was five times stronger in the presence of myristicin, a phenylpropene component that contains methylenedioxyphenyl (MDP), making it a highly effective synergist of xanthotoxin (8-Methoxypsoralen) when used against the corn earworm Heliothis zea (Boddie) (Lepidoptera: Noctuidae) [38]. In this way, furanocoumarins are an effective chemical defense of the chemical diversity necessary for plant defense, e.g., xanthotoxin [39]. When linear xanthotoxin, which is present in many plants of the families Rutaceae and Umbelliferae, was administered to larvae of the southern worm Spodoptera eridania (Stoll) (Lepidoptera: Noctuidae), it showed toxic properties. Its biosynthetic precursor, umbelliferone, lacks this toxicity, and the mechanism of DNA photoinactivation by furanocoumarins is ultraviolet-catalyzed strand cross-linking. Therefore, the ability of a plant to convert umbelliferone to linear furanocoumarins appears to confer broader protection against herbivorous insects [40]. Like most other coumarins, the linear furanocoumarins are photoactivated plant biosynthetic compounds. Ultraviolet A radiation is certainly a major factor determining the toxicity of linear furanocoumarins [41]. The furocoumarins are naturally synthesized with a furan ring in different plant species. The furocoumarin class occurs naturally in various plants, e.g., lemons, limes, and parsnips [42], and now the yellow cedar Ficus petiolaris. Given their growth-inhibiting and feeding-deterrent properties, as well as their insecticidal effect, both the EEFP extract of Ficus petiolaris and its pure allelochemical 8-Methoxypsoralen have the potential to be used as alternative crop protectants against larvae of the insect pest Spodoptera frugiperda.
5. Conclusions
The results of this study show that the ethanolic extract of Ficus petiolaris and its compound 8-Methoxypsoralen has deterrent (larval and pupal weight inhibition) and toxic effects on Spodoptera frugiperda.
Conceptualization, R.F.-B., D.O.S.-S. and J.M.R.-G.; Methodology, R.F.-B., D.O.S.-S., M.Á.R.-L., J.M.R.-G., M.G.V.-C. and O.S.-C.; Validation, R.F.-B., D.O.S.-S. and J.M.R.-G.; Investigation, D.O.S.-S., J.M.R.-G., R.F.-B., M.G.V.-C. and D.A.-M.; Resources, R.F.-B., D.O.S.-S. and J.M.R.-G.; Data curation, R.F.-B., D.O.S.-S. and D.A.-M.; Supervision, R.F.-B., D.O.S.-S., J.M.R.-G., D.A.-M., C.S.-L. and M.Á.R.-L.; Writing—original draft preparation, R.F.-B. and D.O.S.-S.; Writing—review and editing, R.F.-B., D.O.S.-S., J.M.R.-G., D.A.-M. and C.S.-L. Funding acquisition, R.F.-B. All authors have read and agreed to the published version of the manuscript.
The original contributions presented in this study are included in the article/
The authors are grateful to Laboratorio de Fitoquímica y Productos Naturales del Centro de Investigación en Biodiversidad y Conservación-UAEM and the Laboratorio de Entomología del Centro de Desarrollo de Productos Bióticos-Instituto Politécnico Nacional, for the use of their instrumental equipment. We thank Gabriel Flores Franco for the taxonomic identification of the plant species. The authors would like to thank the Centro de Investigaciones Químicas-Universidad Autónoma del Estado de Morelos for the spectroscopic analyses.
The authors declare no conflict of interest.
Footnotes
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Figure 2. Mean weight (mg) of Spodoptera frugiperda fed an artificial diet with EEFP at various concentrations, compared to artificial diet alone. (A) Larval weight (F(2, 6) = 646.67 p ≤ 0.001). (B) Pupal weight (F(2, 6) = 51.72, p ≤ 0.001). Groups with different letters and different color bars are statistically significant different (p ≤ 0.05). Error bars show the standard deviation.
Figure 3. Mean cumulative mortality (±SD) of S. frugiperda with artificial diet with EEFP at various concentrations, compared to controls. Different letters and different bar colors indicate statistically significant different means among treatments (F(2, 6) = 287.96, p [less than] 0.001). LC50 = 1396.9 and LC90 = 3744.4.
Figure 4. Mean mortality larval stages (±SD) of S. frugiperda fed an artificial diet with 8-Methoxypsoralen at various concentrations, compared to controls. Treatments with different letters and bar colors are statistically significant different from each other (F(2, 6) = 601.76; p ≤ 0.001). LC50 = 67.68 ppm and LC90 = 505.17 ppm.
Insectistatic and insecticidal effects of Ficus petiolaris extract in artificial diet against Spodoptera frugiperda.
Treatments | Percentage of Mortality (±SD) | LC50 | LC90 | ||
---|---|---|---|---|---|
Larval 1 | Pupal 2 | Cumulative 3 | (ppm) | ||
500 | 27 ± 6 c | 5 ± 3 bc | 32 ± 6 d | 1396.9 | 3744.4 |
1000 | 30 ± 5 c | 5 ± 2 bc | 35 ± 7 d | ||
1500 | 53 ± 4 b | 7 ± 2 b | 60 ± 6 c | ||
2000 | 56 ± 5 b | 8 ± 3 b | 64 ± 8 c | ||
2500 | 60 ± 5 b | 20 ± 4 a | 80 ± 9 b | ||
Methyl parathion | 100 ± 0 a | ----- | 100 ± 0 a | ||
Artificial diet | 5 ± 1 d | 2 ± 1 c | 7 ± 2 e |
Data based on mean (n = 90). Average values (±SD) followed by the same letter are not statistically significant different (Tukey’s test, p < 0.05). 1 Larval mortality: F(2, 6) = 616.74, p = 0.001. 2 Pupal mortality: F(2, 6) = 24.73, p < 0.001. 3 Cumulative mortality: F(2, 6) = 287.96, p ≤ 0.001. LC50 = 50% lethal concentration; LC90 = 90% lethal concentration, calculated using cumulative mortality.
Insectistactic and insecticidal effects of the fractions of the ethanolic extract of Ficus petiolaris (±SD) on Spodoptera frugiperda.
Treatments (ppm) | Larval Weight 1 | Pupal Weight 2 | Cumulative Mortality 3 | LC50 | LC90 |
---|---|---|---|---|---|
(ppm) | |||||
FpR1 | |||||
1250 | 40.35 ± 2.4 bc | 135.83 ± 1.9 cd | 90 ± 2 b | 857.8 | 2057.9 |
1000 | 42.53 ± 4.5 ab | 138.62 ± 3.1 cd | 83 ± 3 c | ||
750 | 44.81 ± 2.1 ab | 140.82 ± 5.4 bc | 80 ± 4 cd | ||
500 | 45.40 ± 1.2 a | 148.37 ± 2.5 ab | 57 ± 2 e | ||
250 | 46.20 ± 2.7 a | 149.73 ± 1.7 a | 53 ± 4 e | ||
FpR2 | |||||
1250 | 30.35 ± 1.7 d | 136.73 ± 5.2 cd | 100 ± 0 a | 668.46 | 1138.46 |
1000 | 33.53 ± 5.2 cd | 137.25 ± 5.2 cd | 93 ± 5 b | ||
750 | 36.81 ± 2.1 c | 139.72 ± 3.5 bc | 75 ± 3 d | ||
500 | 40.40 ± 2.4 bc | 144.61 ± 1.2 b | 70 ± 5 d | ||
250 | 42.20 ± 6.7 ab | 145.93 ± 2.3 ab | 69 ± 5 d | ||
FpR3 | |||||
1250 | 39.35 ± 3.4 bc | 139.56 ± 2.0 c | 40 ± 5 fg | 1454.2 | 2895.7 |
1000 | 40.25 ± 2.8 bc | 144.68 ± 3.5 b | 30 ± 3 h | ||
750 | 40.81 ± 2.1 bc | 145.30 ± 2.9 ab | 30 ± 2 h | ||
500 | 41.53 ± 1.2 b | 147.74 ± 3.6 ab | 15 ± 5 i | ||
250 | 42.73 ± 2.3 ab | 147.83 ± 3.2 ab | 11 ± 4 j | ||
FpR4 | |||||
1250 | 28.36 ± 2.6 d | 122.56 ± 3.0 e | 72 ± 5 d | 944.79 | 1353.25 |
1000 | 31.47 ± 1.8 d | 126.68 ± 5.6 e | 46 ± 3 f | ||
750 | 33.50 ± 3.4 cd | 131.82 ± 3.7 de | 32 ± 2 h | ||
500 | 35.47 ± 3.3 cd | 134.74 ± 1.6 d | 23 ± 3 i | ||
250 | 36.31 ± 2.1 c | 135.26 ± 4.2 cd | 17 ± 4 i | ||
FpR5 | |||||
1250 | 33.51 ± 6.2 cd | 136.24 ± 1.9 cd | 57 ± 5 e | 931.1 | 2120.8 |
1000 | 35.24 ± 3.1 cd | 137.48 ± 3.1 cd | 42 ± 3 fg | ||
750 | 36.01 ± 2.7 c | 139.50 ± 3.4 c | 39 ± 2 g | ||
500 | 38.11 ± 1.9 c | 144.21 ± 2.5 b | 31 ± 3 h | ||
250 | 39.05 ± 1.0 c | 144.57 ± 1.7 b | 19 ± 4 i | ||
Methyl parathion | ----- | ----- | 100 ± 0 a | ||
Artificial diet | 43.73 ± 2.3 ab | 146.05 ± 2.4 ab | 5 ± 3 j |
Data based on mean (n = 90). Average values (±SD) followed by the same letter are not statistically significant different (Tukey’s test, p < 0.05). 1 Weight larval: F(2, 26) = 68.02; p = 0.001. 2 Weight pupae: F(2, 26) = 58.89; p = 0.001. 3 Cumulative mortality: F(2, 26) = 871.24; p ≤ 0.001. LC50 = 50% lethal concentration; LC90 = 90% lethal concentration, calculated using cumulative mortality.
Insectistactic and insecticidal effects of 8-Methoxypsoralen of the ethanolic extract of Ficus petiolaris (±SD) on Spodoptera frugiperda.
8-Methoxypsoralen (ppm) | Larval Weight 1 | Pupal Weight 2 | Larval Mortality 3 | LC50 | LC90 |
---|---|---|---|---|---|
(ppm) | |||||
100 | 15.34 ± 2.9 c | 112.55 ± 4.8 d | 100 ± 0 a | 67.68 | 505.17 |
75 | 16.17 ± 3.8 c | 124.83 ± 5.7 c | 96 ± 3 b | ||
50 | 25.24 ± 5.1 b | 126.26 ± 2.9 c | 88 ± 4 c | ||
25 | 28.96 ± 4.4 b | 136.93 ± 3.6 b | 81 ± 4 d | ||
12.5 | 31.35 ± 2.3 b | 137.48 ± 2.5 b | 64 ± 3 e | ||
Methyl parathion | ----- | ----- | 100 ± 0 a | ||
Artificial diet | 47.53 ± 3.7 a | 148.25 ± 2.4 a | 8 ± 2 f |
Data based on mean (N = 30). Average values (±SD) followed by the same letter are not statistically significant different (Tukey’s test, p < 0.05). 1 Larval weight: F(2, 6) = 239.96; p ≤ 0.001. 2 Pupal weight: (2, 6) = 310.22; p = 0.001. 3 Larval mortality: F(2, 6) = 601.76; p ≤ 0.001. LC50 = 67.68 ppm and LC90 = 505.17 ppm.
Supplementary Materials
The following supporting information can be downloaded at:
References
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Abstract
In this study, we tested the insectistatic and insecticidal effects of the ethanolic extract of stems and bark of Ficus petiolaris Kunth (Moraceae) in laboratory bioassays with an artificial diet against newly hatched larvae of the fall armyworm, Spodoptera frugiperda Smith (Lepidoptera: Noctuidae). The extract was evaluated at five different concentrations (500, 1000, 1500, 2000, and 2500 ppm). The 2500 ppm extract had the strongest inhibitory effects on larval (89%) and pupal (20%) weight as well as the highest mortality (80%). The positive control, methyl parathion (Methyl Parathion®) at 1%, eliminated 100% of the fall armyworm, and the negative control (artificial diet) had a mortality of only 5%. Chemical fractionation of F. petiolaris stem and bark extract produced five fractions of FpR1-5, each of which was evaluated at 250, 500, 750, 1000, and 1250 ppm. FpR4 presented the strongest inhibitory effect, reducing the weight of the larva and pupa by 35% and 18%, while FpR2 had the strongest insecticidal effect, with 90% mortality at 1250 ppm. The pure 8-Methoxypsoralen compound extracted from this fraction was even more effective, with 100% S. frugiperda mortality at 100 ppm. The 50% lethal concentration (LC50) of 8-Methoxypsoralen was 67.68 ppm. Our results indicate that the F. petiolaris extract showed toxic activity against the fall armyworm, and its compound 8-Methoxypsoralen showed strong insecticidal activity at low concentrations.
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1 Departamento de Interacciones Planta-Insecto, Centro de Desarrollo de Productos Bióticos, Instituto Politécnico Nacional, Carretera Yautepec-Jojutla Km 6, Col. San Isidro, Yautepec C.P. 62731, Morelos, Mexico;
2 Escuela de Estudios Superiores del Jicarero, Universidad Autónoma del Estado de Morelos, Carretera Galeana-Tequesquitengo s/n, Jojutla C.P. 62907, Morelos, Mexico;
3 Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Guerrero, Av. Lázaro Cárdenas s/n, Ciudad Universitaria Sur, Chilpancingo C.P. 39000, Guerrero, Mexico;
4 Facultad de Química, Universidad Autónoma de Querétaro, Cerro de las Campanas s/n, Col. Las Campanas, Santiago de Querétaro C.P. 76010, Querétaro, Mexico;
5 Facultad de Ciencias Biológicas, Universidad Autónoma del Estado de Morelos, Av. Universidad No. 1001, Col. Chamilpa, Cuernavaca C.P. 62209, Morelos, Mexico;
6 Facultad de Ciencias Químicas e Ingeniería, Universidad Autónoma del Estado de Morelos, Av. Universidad No. 1001, Col. Chamilpa, Cuernavaca C.P. 62209, Morelos, Mexico;
7 Escuela de Estudios Superiores del Jicarero, Universidad Autónoma del Estado de Morelos, Carretera Galeana-Tequesquitengo s/n, Jojutla C.P. 62907, Morelos, Mexico;