Plutella Xylostella Larvae Research Articles

Characterization of larval gut microbiota of two endoparasitoid wasps associated with their common host, Plutella xylostella (Linnaeus) (Lepidoptera: Plutellidae).

Insect gut microbes play important roles in digestion, metabolism, development, and environmental adaptation. Parasitoid wasps are one of the most important biological control agents in pest control, while the gut microbial species compositions and the associated functions have been poorly investigated. Two endoparasitoid wasps, Cotesia vestalis and Diadromus collaris, parasitize the larval stage and pupal stage of the diamondback moth, Plutella xylostella, respectively. Using whole-genome shotgun metagenomic sequencing, we characterized the gut microbial composition, diversity, and potential functional roles associated with the two parasitoid wasp larvae. The results reveal that Proteobacteria and Firmicutes are the dominant phyla in the gut of C. vestalis and D. collaris larvae, with Rhizobium and Enterococcus being the dominant genera. The putative microbial functions associated with the two parasitoid wasps might play a virtual role in assisting in consuming the host's nutritional composition. The enriched CAZymes family genes are primarily involved in the degradation and synthesis of chitin. Despite the richness of microbial species and communities, the microbes species and the microbial community structure exhibit significant similarity between the two parasitoid wasps and between the parasitoid wasp and the host P. xylostella. Notably, the prevalence of the genus Enterococcus shared among them suggests a possible link of gut microbes between the host and their associated parasitoids. Our study offers insights into the gut microbe-based interactions between the host and parasitoid wasps for the first time, potentially paving the way for the development of an ecologically friendly biocontrol strategy against the pest P. xylostella.IMPORTANCEEndoparasitoid wasps spend the majority of their lifespan within their host and heavily rely on the host's nutrition for survival. There is limited understanding regarding the composition and physiological impacts of gut microbial communities in parasitoid wasps, particularly during the larval stage, which is directly linked to the host. Based on a thorough characterization of the gut microbe and comprehensive comparative analysis, we found the microbial species of the larval parasitoid wasp Cotesia vestalis and the pupal parasitoid wasp Diadromus collaris were similar, sharing 159 genera and 277 species, as were the microbial community structure. Certain of the dominant microbial strains of the two parasitoid wasps were similar to that of their host Plutella xylostella larvae, revealing host insect may affect the microbial community of the parasitoid wasps. The putative microbial functions associated with the parasitoid wasp larvae play an important role in dietary consumption.

Redox-Responsive Nanopesticides Based on Natural Polymers for Environmentally Safe Delivery of Pesticides with Enhanced Foliar Dispersion and Washout Resistance.

Based on the modified cross-linking of the degradable natural polymers chitosan oligosaccharides (COS) and gelatin (GEL) via introduction of a functional bridge 3,3'-dithiodipropionic acid, this study constructed an environmentally responsive dinotefuran (DNF) delivery system (DNF@COS-SS-GEL). The introduction of the disulfide bond (-S-S-) endowed DNF@COS-SS-GEL with redox-responsive properties, allowing for the rapid release of pesticides when stimulated by glutathione (GSH) in the simulated insect. Compared with commercial DNF suspension concentrate (DNF-SC), DNF@COS-SS-GEL showed superior wet spreading and retention performance on cabbage leaves with a reduced contact angle (57°) at 180 s and 4-fold increased retention capacity after rainfall washout. Nanoencapsulation effectively improved the UV-photostability with only a 31.4% decomposition rate of DNF@COS-SS-GEL at 96 h. The small scale and large specific surface area resulted in excellent uptake and transportation properties in plants as well as higher bioactivity against Plutella xylostella larvae. This study will help promote sustainable agricultural development by reducing environmental pollution through improved pesticide utilization.

Synergism of Cry1 Toxins by a Fusion Protein Derived from a Cadherin Fragment and an Antibody Peptide.

Synergistic factors can enhance the toxicity of Bt toxins and delay the development of Bt resistance. Previous research has demonstrated that a Helicoverpa armigera cadherin fragment (HaCad-TBR) increased the toxicity of Cry1Ac in Plutella xylostella larvae but did not have a synergistic effect on Cry1B, Cry1C, and Cry1F toxins. In this study, a fusion protein (HaCad-TBR-2D3 VL) derived from HaCad-TBR and a Bt Cry1-specific antibody peptide was expressed in Escherichia coli. The HaCad-TBR-2D3 VL enhanced Cry1Ac toxicity more efficiently in insects and Sf9 cells than HaCad-TBR and also significantly increased the toxicity of Cry1B, Cry1C, and Cry1F toxins in insects. Further investigation indicated that the improved stability in insect midguts and higher binding capacity with Bt toxins contributed to the enhanced synergism of HaCad-TBR-2D3 VL over HaCad-TBR. This study suggested that Bt antibody fragments can potentially broaden the synergistic range of Bt receptor fragments, providing a theoretical foundation for developing broad-spectrum synergists for other biopesticides.

Potential of secondary metabolites from soil-derived actinomycetes as juvenile hormone disruptor and insecticides

Actinomycetes represent sources of potential alternatives to chemicals as biological pest control agents, and several actinomycetes are known to produce metabolites with insecticidal activities against insect pests. Although these metabolites are eco-friendly and effective, it is necessary to identify a novel insect-specific insecticidal agents that can minimize the occurrence of resistance. Thus, in this study, we screened the juvenile hormone disruptor (JHD) and insecticidal activities of crude acetone extracts derived from various actinomycetes as novel biopesticide candidates. A total of 300 soil-derived actinomycetes were isolated from various sites in Korea, and were screened for their JHD and insecticidal activities against a range of important medical and agricultural pests. Among them, eight extracts showed high levels of JHD activity. The AN120590 strain showed a broad insecticidal spectrum with stably maintained JHD properties. Additionally, the dead larvae of Aedes albopictus and Plutella xylostella treated with acetone extracts of the AN120590 strain displayed morphological deformities similar to those of insects treated with juvenile hormone-based insect growth regulators (IGRs). These findings indicate that actinomycetes represent promising sources of novel IGRs that can be effectively applied as insecticides with a broad insecticidal spectrum.

Minimizing IP issues associated with gene constructs encoding the Bt toxin - a case study

BackgroundAs part of a publicly funded initiative to develop genetically engineered Brassicas (cabbage, cauliflower, and canola) expressing Bacillus thuringiensis Crystal (Cry)-encoded insecticidal (Bt) toxin for Indian and Australian farmers, we designed several constructs that drive high-level expression of modified Cry1B and Cry1C genes (referred to as Cry1BM and Cry1CM; with M indicating modified). The two main motivations for modifying the DNA sequences of these genes were to minimise any licensing cost associated with the commercial cultivation of transgenic crop plants expressing CryM genes, and to remove or alter sequences that might adversely affect their activity in plants.ResultsTo assess the insecticidal efficacy of the Cry1BM/Cry1CM genes, constructs were introduced into the model Brassica Arabidopsis thaliana in which Cry1BM/Cry1CM expression was directed from either single (S4/S7) or double (S4S4/S7S7) subterranean clover stunt virus (SCSV) promoters. The resulting transgenic plants displayed a high-level of Cry1BM/Cry1CM expression. Protein accumulation for Cry1CM ranged from 5.18 to 176.88 µg Cry1CM/g dry weight of leaves. Contrary to previous work on stunt promoters, we found no correlation between the use of either single or double stunt promoters and the expression levels of Cry1BM/Cry1CM genes, with a similar range of Cry1CM transcript abundance and protein content observed from both constructs. First instar Diamondback moth (Plutella xylostella) larvae fed on transgenic Arabidopsis leaves expressing the Cry1BM/Cry1CM genes showed 100% mortality, with a mean leaf damage score on a scale of zero to five of 0.125 for transgenic leaves and 4.2 for wild-type leaves.ConclusionsOur work indicates that the modified Cry1 genes are suitable for the development of insect resistant GM crops. Except for the PAT gene in the USA, our assessment of the intellectual property landscape of components presents within the constructs described here suggest that they can be used without the need for further licensing. This has the capacity to significantly reduce the cost of developing and using these Cry1M genes in GM crop plants in the future.

A flexible modulated pesticide release platform through poly(urethane-urea) microcapsules: effect of different crosslinkers compositions.

Polymeric microcapsules (MCs) have become an important issue and have attracted increasing attention because of their tunable physical and chemical properties. Diverse shell structures can confer multiple properties on MCs. Different polyols (1,4-butanediol and glycerin) and polyamines (triethylenetetramine and isophorondiamine) were selected as crosslinkers to obtain emamectin benzoate (EB)-loaded poly(urethane-urea) MCs (PU-MCs) by interfacial polymerization. The four obtained PU-MCs showed sphericity with different degrees of smoothness on their surfaces, and displayed a uniform size distribution ranging from 500 to 700 nm. Moreover, transmission electron microscopy showed that the shell thickness was roughly uniform, and was greatly influenced by the type and structure of the crosslinker. GI-MCs, prepared using glycerin and isophorondiamine, had the largest shell thickness. GT-MCs, obtained using glycerin and triethylenetetramine, had the highest encapsulation efficiency and drug-loading content, and BT-MCs, obtained using mixtures of 1,4-butanediol and triethylenetetramine, had the fastest release behavior. Thermogravimetric analysis revealed that the greater the degree of shell crosslinking, the higher decomposition temperature and the greater the thermal stability. A BT-MC suspension had the lowest viscosity and contact angle with the best wettability. Bioassay experiments showed that BT-MCs exhibited good insecticidal activity against Plutella xylostella larvae with a half-maximal lethal concentration of 4.19 mg/L. Furthermore, a BT-MC suspension showed good thermal and light stability, with potential applications in minimizing the toxicity of EB through sustained release. Various properties of EB-loaded PU-MCs were modulated through simple selection of different polyols and polyamines during fabrication, which might have an important role in constructing the pesticide delivery system and improving pesticide utilization. © 2024 Society of Chemical Industry.

Effect of Brucea javanica Oil on the Toxicity of β-Cypermethrin Emulsifiable Concentrate Formulation.

Enhancing the performance of traditional pesticide formulations by improving their leaf surface wetting capabilities is a crucial approach for maximizing the pesticide efficiency. This study develops an emulsifiable concentrate (EC) of 4.5% β-cypermethrin containing Brucea javanica oil (BJO). The incorporation of BJO aims to improve the leaf-wetting properties of the EC formulation and enhance its insecticidal effectiveness. The droplet size and emulsion characteristics of β-CYP EC emulsion with varying concentrations of the emulsifier were evaluated, and changes after incorporating BJO were assessed to develop the optimal formulation. A comprehensive comparison was conducted among commercial 4.5% β-cypermethrin EC (β-CYP EC-1), 4.5% β-cypermethrin EC with BJO (β-CYP EC-2), and 4.5% β-cypermethrin EC without BJO (β-CYP EC-3). This comparison encompassed various factors including storage stability, insecticidal activity, cytotoxicity, and wetting performance on cabbage leaves. The results indicated that the ideal emulsifier concentration was 15% emulsifier 0201B. β-CYP EC-2 demonstrated superior wetting properties on cabbage leaves (the wetting performance of β-CYP EC-2 emulsion on cabbage leaves is 2.60 times that of the β-CYP EC-1 emulsion), heightened insecticidal activity against the third larvae of Plutella xylostella [diamondback moth (DBM)] [the insecticidal activity of the β-CYP EC-2 emulsion against the third larvae of DBM is 1.93 times that of the β-CYP EC-1 emulsion (12 h)], and more obvious inhibitory effects on the proliferation of DBM embryo cells than the other tested formulations. These findings have significant implications for advancing pest control strategies and promoting sustainable and effective agricultural practices.

MiR-2b-3p Downregulated PxTrypsin-9 Expression in the Larval Midgut to Decrease Cry1Ac Susceptibility of the Diamondback Moth, Plutella xylostella (L.).

Crystal (Cry) toxins, produced by Bacillus thuringiensis, are widely used as effective biological pesticides in agricultural production. However, insects always quickly evolve adaptations against Cry toxins within a few generations. In this study, we focused on the Cry1Ac protoxin activated by protease. Our results identified PxTrypsin-9 as a trypsin gene that plays a key role in Cry1Ac virulence in Plutella xylostella larvae. In addition, P. xylostella miR-2b-3p, a member of the micoRNA-2 (miR-2) family, was significantly upregulated by Cry1Ac protoxin and targeted to PxTrypsin-9 downregulated its expression. The mRNA level of PxTrypsin-9, regulated by miR-2b-3p, revealed an increased tolerance of P. xylostella larvae to Cry1Ac at the post-transcriptional level. Considering that miR-2b and trypsin genes are widely distributed in various pest species, our study provides the basis for further investigation of the roles of miRNAs in the regulation of the resistance to Cry1Ac and other insecticides.

Evaluation of Insecticidal Performances of Jayanti Plant (Sesbania sesban) for Integrated Control of Cabbage Caterpillar (Plutella xylostella)

The study on the use of local natural materials as a source of insecticides is intended, among other things, to reduce farmers' dependency on using synthetic chemicals. This research was conducted to obtain the most selective insecticidal active ingredient from Sesbania sesban plant for controlling cabbage caterpillars, choose a target mode of action that has a safe impact on the environment, and determine insecticidal stability during storage of the material prior to application and its stability in water after application. The dried powder of S. sesban leaves was extracted in stages using a series of solvents, namely hexane, DCM, ethanol and water, respectively. Each of the extract fractions produced was tested for its lethal toxicity against Plutella xylostella larvae and Diadegma semiclausum imago. Each of mortality data were processed by probit analysis to produce LC50(s) to determine their insecticidal selectivity. Subsequent bioassays were carried out using extract-water of S. sesban leaves and the data were processed by probit analysis to determine their anti-ovipositor, ovicidal powers and anti-feedant properties against P. xylostella. The insecticidal stability of S. sesban was also studied according to variations in the storage time of the simplicia before extraction, the storage time of the extract before dissolution, and its stability in water during application. The data was processed using ANOVA. The results show that the insecticidal activity of the ethanol extract fraction of S. sesban leaves for controlling cabbage caterpillars was very selective, namely very toxic to P. xylostella but very less toxic to D. semiclausum. Against P. xylostella, S. sesban leaf extract has also been proven to significantly prevent oviposition, inhibit egg hatching and feeding activity. The insecticidal power of S. sesban did not decrease significantly during storage of the simplicia for less than three months and during storage of the extract for less than seven days. However, the toxicity of the extract solution decreased drastically after 24 hours of application

Coprecipitation-based synchronous chlorantraniliprole encapsulation with chitosan: carrier-pesticide interactions and release behavior.

Controlled-release pesticide formulations have emerged as a promising approach towards sustainable pest control. Herein, an environment-friendly formulation of insecticide chlorantraniliprole (CAP) was fabricated through a simple approach of coprecipitation-based synchronous encapsulation by chitosan (CTS), with carrier-pesticide interaction mechanism and release behavior investigated. The resulting CAP/CTS controlled-release formulation (CCF) showed a good loading content of 28.1% and a high encapsulation efficiency of 75.6%. Instrument determination in combination with molecular dynamics (MD) simulations displayed that the primary interactions between CAP and CTS were physical adsorption and complicated hydrogen (H)-bonds, which formed dominantly between NH in amides [or nitrogen (N) in ring structures] of CAP and hydroxyl (or amino) groups of CTS, as well as oxygen (O) in CAP with hydrogen in CTS or H2 O molecules. The in vitro release tests exhibited obvious pH/temperature sensitivity, with release dynamics following the first-order or Ritger-Peppas model. As the temperature increased, the CAP release process of the Ritger-Peppas model changed from Case-II to anomalous transport, and ultimately to a Fickian diffusion mechanism. The control effect against Plutella xylostella larvae also was evaluated by toxicity tests, where comparable efficacy of CCF to the commercial suspension concentrate was obtained. The innovative, easy-to-prepare CCF can be used as a formulation with obvious pH/temperature sensitivity and good efficacy on target pests. This work contributes to the development of efficient and safe pesticide delivery systems, especially using the natural polymer materials as carriers. © 2023 Society of Chemical Industry.

A Potential Lead for Insect Growth Regulator: Design, Synthesis, and Biological Activity Evaluation of Novel Hexacyclic Pyrazolamide Derivatives.

Ecdysone receptor (EcR) and chitinase play a critical role in the molting stage of insect pests. Each of them is considered a promising target for the development of novel insect growth regulators (IGRs). In the present paper, a total of 24 (23 novel) hexacyclic pyrazolamide derivatives were designed and synthesized by reducing the heptacycle and inserting small flexible linkers on the basis of the previously discovered dual-target compound D-27 acting simultaneously on EcR and Ostrinia furnacalis chitinase (OfChtI). Their insecticidal activities against Plutella xylostella, Spodoptera frugiperda, and Ostrinia furnacalis larvae were evaluated. The results revealed that the insecticidal activity was not significantly enhanced when the heptacycle on the pyrazole ring was reduced to a hexacycle. However, the insertion of an additional methylene spacer between the substituted phenyl ring and the amide bond can improve the insecticidal activity. Among the derivatives, the most potent compound, 6j, exhibited promising insecticidal activities against P. xylostella and S. frugiperda. Further protein binding assays and molecular docking indicated that 6j could target both EcR and OfChtI, and is a potential lead compound for IGRs. The present work provides valuable clues for the development of new dual-target IGRs.

Stability of Anti-Insect Ingredient from Jayanti Plants (Sesbania sesban) for Integrated Control of Cabbage Pest

The specific purpose of this study was to determine the stability of S. sesban insecticide during storage before application and its stability in water during application. The dry powder of S. sesban leaves was extracted using water as the single solvent. Bioassay of S. sesban leaf extract was carried out on cabbage caterpillars (Plutella xylostella larvae) and Diadegma semiclausum imago on cabbage plants using a variation of storage time design. Mortality data of P. xylostella larvae and D. semiclausum imago were respectively processed by probit analysis to determine LC50. The results showed that the insect repellent content of S. sesban leaves was a compound from the saponin group and was unstable during simplicia storage, extract storage and also unstable in water before and during application. Storage of S. sesban leaf simplicia from 1 to 3 months only slightly reduced the lethal toxicity of the extract to P. xylostella larvae (mortality from 95 to 80% or LC50 from 28.82 to 28.83 ppm), but after 6 to 12 months storage, the lethal toxicity decreased drastically (mortality was 12.5 to 1.25% and LC50 was 247.99 ppm until calculated). Storage of S. sesban leaf extract from 7 to 15 days had resulted in a sharp decrease in lethal toxicity to P. xylostella larvae (mortality 70 to 40% and LC50 34.05 to 59.43 ppm) and 30 days storage causes the insect repellent to be inactive. (mortality was only 1.25% and LC50 was unaccounted for). Exposure to a solution of S. sesban leaf extract for 24 to 48 hours caused a decrease in lethal toxicity to P. xylostella larvae (mortality 32 to 28% and LC50 62.63 to 64.85 ppm) and after the solution was stored for 72 hours, the insect repellent was almost no active again (mortality was only 1.25 and LC50 was unstoppable). All storage treatments of insect repellent from S. sesban leaves showed no significant difference in the effect of lethal toxicity on D. semiclausum imago. In all the results of the bioassays, the mortality of D. semiclasum was 0 to 1.25% and each of all LC50(s) was unaccounted for

Identification, Pathogenicity, and Culture Conditions of a New Isolate of Cordyceps javanica (Hypocreales: Cordycipitaceae) From Soil.

This study decribes a highly effective insecticidal isolate of Cordyceps javanica (Frieder. & Bally) (Hypocreales: Cordycipitaceae) named IJ-tg19, which was isolated from soil. Spray bioassays were performed with IJ-tg19 on Myzus persicae (Sulzer) (Hemiptera: Aphididae) adults, third-instar nymphs of Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae), and third-instar larvae of Plutella xylostella (Linnaeus) (Lepidoptera: Plutellidae) to determine the pathogenicity of the isolate. The corrected mortality rates for all three pests were 100% when the conidia concentration was 1 × 106 conidia/ml, the lowest concentration in this study, and the median survival times (MST) were 4, 4, and 3 d. The MST shortens with increasing conidia concentration. The effects of laboratory culture conditions on the sporulation and growth of the isolate were also studied. This isolate had the greatest conidia production and fastest growth rate on malt extract agar medium at 25°C. The amount of conidia produced had positive correlation to light duration, with the highest production at 24 hr light. The growth of mycelium can adapt to a moderately alkaline environment, but the optimum conidial production occurred at the pH of 7. Our finding and research will be useful in biocontrol programs that are considering using the new isolate of C. javanica against greenhouse pests.

Design, synthesis, and biological activity of novel spiro-pyrazolo[1,5-a]quinazolines derivatives as potential insecticides.

Arylpyrazole insecticides display broad-spectrum insecticidal activity against insect pests. However, the high toxicity toward honeybees associated with fipronil prohibits its agronomic utility. To explore reducing the toxicity of aryl pyrazole analogs to bees, a series of new spiro-pyrazolo[1,5-a]quinazoline derivatives were designed and synthesized. Bioassay results showed that these compounds exhibited good insecticidal activity. In particular, the insecticidal activity of compound 5f against Plutella xylostella larvae (median lethal contentration, LC50 =1.43 mg L-1 ) was equivalent to that of fipronil. Moreover, some compounds also showed good insecticidal activity against Solenopsis invicta. Importantly, the bee toxicity study confirmed that compound 5f had much lower acute oral toxicity, with a median lethal dose (LD50 ) = 1.15 μg bee-1 that was three to four orders of magnitude greater than that of fipronil (0.0012 μg bee-1 ). Electrophysiological studies were conducted using honeybee γ-aminobutyric acid receptor heterologously expressed in Xenopus oocytes to explain the reduced bee toxicity of compound 5f. The inhibitory effect of compound 5f (16.29 μmol L-1 ) was determined to be approximately 700-fold lower than that of fipronil (0.023 μmol L-1 ). These spiro-pyrazolo[1,5-a]quinazoline derivatives could be potential candidates and lead structures for the discovery of novel insecticides with low bee toxicity. © 2022 Society of Chemical Industry.

Insecticidal Activities and GC-MS Analysis of the Selected Family Members of Meliaceae Used Traditionally as Insecticides.

The environmental and health risks associated with synthetic pesticides have increased the demand for botanical insecticides as safer and biodegradable alternatives to control insect pests in agriculture. Hence in this study, five Meliaceae species were evaluated for their insecticidal activities against the Spodoptera frugiperda and the Plutella xylostella larvae, as well as their chemical constituents. Repellence, feeding deterrence, and topical application bioassays were employed to evaluate their insecticidal activities. GC-MS analysis was performed to identify chemical compounds present in each plant. The repellence bioassay indicated that Melia azedarach extracts exhibited the highest repellence percentage against S. frugiperda (95%) and P. xylostella (90%). The feeding deterrence bioassay showed that M. azedarach and Trichilia dregeana extracts displayed excellent antifeeding activity against the S. frugiperda (deterrent coefficient, 83.95) and P. xylostella (deterrent coefficient, 112.25), respectively. The topical application bioassay demonstrated that Ekebergia capensis extracts had the highest larval mortality against S. frugiperda (LD50 0.14 mg/kg). Conversely, M. azedarach extracts showed the highest larval mortality against P. xylostella (LD50 0.14 mg/kg). GC-MS analysis revealed that all plant extracts had compounds belonging to the two noteworthy groups (phenols and terpenes), which possess insecticidal properties. Overall, this study lends scientific credence to the folkloric use of Meliaceae species as potential biocontrol agents against insect pests.

Laboratory Evaluation of the Insecticidal Toxins from Entomopathogenic Nematode Symbiotic Bacteria to Control Vegetable Diseases and Pests

Aims: Entomopathogenic nematode (EPN) and its symbiotic bacterium are used worldwide as microbial control agents. Toxins from EPN symbiotic bacteria were isolated and provided basis for using this potential resource as biocontrol agent against vegetable diseases and pests.
 Study Design: The toxins were extracted from 28 strains of bacteria associated with entomopathogenic nematodes. The insecticidal activity and antibiotic activity against vegetable diseases and pests were determined through bioassay.
 Place and Duration of Study: College of Bioscience and Biotechnology, between May 2020 and September 2021.
 Methodology: The toxins were extracted by (NH4)2SO4 precipitation method. The insecticidal activities and antibiotic activities were evaluated using bioassay in the laboratory.
 Results: The toxins of the symbiotic bacteria associated with EPN had certain insecticidal activities on the first instar larvae of Plutella xylostella and Laphygma exigua, and strain SY5 showed the most obvious antifungal activities against Trichothecium roseum and Fusarium oxysporum.
 Conclusions: The toxins of the EPN symbiotic bacteria SY5 had good insecticidal activity and antibiotic activity. Therefore, it has the potential for use against vegetable diseases and pests as biocontrol agents.

Anti-Ovipositor Power of Jayanti Plant (Sesbania sesban) for Integrated Control of Cabbage Pest (Plutella xylostella)

Cabbage harvest failure is often caused by failure to control cabbage pests. Cabbage pest control by eradicating Plutella xylostella larvae is considered less effective because the cabbage plant has been damaged due to infection with P. xylostella eggs and larvae. Thus, the control target needs to be shifted to prevent P. xylostella amago from laying eggs (ova-positioning) on cabbage plants. This study aims to determine the anti-ovipositor power of Sesbania sesban leaf extract against P. xylostella. The leaves of S. sesban were air-dried and then ground. The dry powder (simplicia) of S. sesban leaves was extracted with water as the sole solvent. The water-extract of S. sesban leaves was tested for anti-ovipositor against P. xylostella in situ on cabbage plants according to a completely randomized block design. Experimental groups were made according to variations in extract concentration. Complete randomization was carried out on experimental units in each group according to the variation in the length of exposure to the extract before access to the ovipositor was opened (Cages were opened after 1,2,3,4,5 and 6 days from the time of extract spraying). Each experimental unit consisted of 4 cabbage plants as replicates. The treatment was given by spraying a solution of S. sesban leaf extract with a concentration according to the group evenly on the entire leaf surface of each cabbage plant. The observed research variable was the number of P. xylostella eggs per cabbage plant after the cage of cabbage plant unit was opened. The results showed that the leaf extract of S. sesban could inhibit P. xylostella as an ovipositor to lay its eggs on cabbage plants effectively for up to three days. With concentrations of 1.83 to 2.14 ppm, the aqueous extract of S. sesban leaves did not inhibit P. xylostella egg laying, but with concentrations of 478.63 to 1283.88 ppm it could inhibit 50 to 100%.

A possible mechanism of Cry7Ab4 protein in delaying pupation of Plutella xylostella larvae.

Cry toxins produced by Bacillus thuringiensis (Bt) are well known for their insecticidal activities against Lepidopteran, Dipteran, and Coleopteran species. In our previous work, we showed that trypsin-digested full-length Cry7Ab4 protoxin did not have insecticidal activity against Plutella xylostella larvae but strongly inhibited their growth. In this paper, we expressed and purified recombinant active Cry7Ab4 toxic core from Escherichia coli for bioassay and identified its binding proteins. Interestingly, Cry7Ab4 toxic core exhibited activity to delay the pupation of P. xylostella larvae. Using protein pull-down assay, several proteins, including basic juvenile hormone-suppressible protein 1-like (BJSP-1), were identified from the midgut juice of P. xylostella larvae as putative Cry7Ab4-binding proteins. We showed that feeding P. xylostella larval Cry7Ab4 toxic core upregulated the level of BJSP-1 mRNA in the hemocytes and fat body and decreased the free juvenile hormone (JH) level in larvae. BJSP-1 interacted with Cry7Ab4 and bound to free JH in vitro. A possible mechanism of Cry7Ab4 in delaying the pupation of P. xylostella larvae was proposed.

Biological, morphological, and molecular characterization of the baculovirus PlxyMNPV_LBIV-11, and its virulence towards Plutella xylostella, Trichoplusia ni, and Spodoptera frugiperda larvae.

PlxyMNPV_LBIV-11 is an alphabaculovirus strain, isolated from Plutella xylostella larvae. This work characterized this strain at a biological, morphological, and molecular level to evaluate its similarity with other baculoviruses. Its ultrastructure showed a multiple arrangement of nucleocapsids within enveloped virions, all occluded within large cubical polyhedra. PlxyMNPV_LBIV-11 showed infectivity on the Hi5 and Sf9 cell lines, despite these being from heterologous origin. This in vitro infectivity was observed using either BVs or by transfection with genomic DNA. Restriction fragment patterns of PlxyMNPV_LBIV-11, using the enzymes EcoRI, BamHI and HindIII, showed a high relationship with those patterns shown by AcMNPV, except for one or two differential bands with each enzyme. Sequences of core genes lef-8 and lef-9 and the conserved polh gene showed identities ranging from 98 to 100% when compared with those of AcMNPV. Somewhat lower was the sequence identity of the gp64 gene (94%) as compared with those of AcMNPV and PlxyMNPV_CL3, which might be related to the difference in virulence. Besides, the presence of this gene in PlxyMNPV_LBIV-11 indicates that it belongs to group 1 of alphabaculoviruses. A phylogram was estimated with the core and conserved gene sequences, corroborating its high relationship with AcMNPV and PlxyMNPV_CL3. Bioassays were performed with P. xylostella larvae reared on a meridic diet, whose LC50 values indicated lower virulence than AcMNPV when tested against P. xylostella, Spodoptera frugiperda, and Trichoplusia ni larvae. Its virulence against S. frugiperda was only seven times lower than AcMNPV. Its potential as a biological control agent is discussed.

Effect of Biocidal Substances on Population Dynamics of Plutella Xylostella Linnaeus, 1758, Auxiliary Insects, and Cabbage Production in the Suburban Area of Dakar

This study was conducted to determine the effects of biopesticides, used in the control of Plutella xylostella, major bioaggressor of cabbage, on its auxiliaries as well as on cabbage quality. It took place in a field of 65 m2, located in Malika, peri-urban area of Dakar. Treatments, based on Crateva religiosa (200g/l), Calotropis procera (400g/l), a mixture of the two products, and the control, were performed every two weeks on a random block device. Analysis of the results was done with the Rstudio software. Results showed that untreated plots were more infested with Plutella xylostella larvae (p-value = 0.01271). Efficacy of Calotropis procera on P. xylostella larvae has been demonstrated. The effect was all the more interesting since auxiliaries were not affected by treatments, as the regular sampling revealed. A satisfactory agronomic result was obtained at the end of this test, with as indicator, a much larger harvest from the treated plots.