Research Progress of Main Pests in Sweetsop ( Annona squamosa L. ) and Its Insecticide Activity

The research progress on pests and insecticide activity of Annona squamosa L. is summarized in the paper, including symptom, occurrence regularity and control methods. Furthermore, the comprehensive technique for controlling pests of A. squamosa in the future is preliminarily discussed.

Insecticidal Activity and Field Efficacy of Novel Insecticide Cyhalodiamide against Different Lepidoptera Pests

[ Objective] The paper was to study the insecticidal activity and field efficacy of cyhalediamide, a novel insecticide developed by Zhejiang Research Institute of Chemical Industry, Ltd. [ Method] Toxicity of cyhalodiamide against Plutella xylostella, Spodoptera exigua, Prodenia litura and Chilo suppressalis were tasted in the laboratory by leaf-dipping method, and the field efficacy of cyhalodiamide against P. xylostella, S. exigua, P. litura, Pieris rapae, C. suppressalis and Cnaphalocrocis medinalis was evaluated. [ Result] The LCs0 values of cyhalodiamide against P. xylostella, S. exigua, P. litura and C. suppressalis were 0. 046, 0.072, 0.071 and 1.95 mg/L, respectively, almost equivalent to that of flubendiamide and ehlorantraniliprole. The control efficacy of 20% Z4042 SC （30 g/hm2） against P. xylostella, S. exigua, P. litura and P. rapae was 90.1% - 96.3 % after spraying for 7 d, and its control efficacy against C. suppressalis and C. medinalis was 82.9% -90.5% after spraying for 15 d. No crop injury was observed at the test dosage. [Conclusion] The study provided reference for field control against Lepidoptera pests.

Synthesis and Insecticidal Activity of Derivants of 3,13-Ester Mesaconitine

[Objective] The experiment aimed to explore the relations between structure of aconitine, aconitine derivatives and insecticide. [Method] The aconitine derivatives such as 3-acetyl mesaconitine, 3-propionyl mesaconitine, 3-acetyl-13-Benzoyl-mesaconitine and 3-propionyl-13-Benzoyl-mesaconitine were synthesized by mesaconitine with acetic anhydride or propionic anhydride and their insecticidal activities were also determined. [Result] When the concentration was 500 mg/L, the insecticidal activities of mesaconitine against Nilaparvata legen and Aphis were 50% and 30% respectively while the insecticidal activities of 3-acetyl mesaconitine, 3-propionyl mesaconitine, 3-acetyl-13-Benzoyl-mesaconitine and 3-propionyl-13-Benzoyl-mesaconitine against Aphis medicagini were 40%, 30%, 30% and 20% respectively at 500 mg/L. [Conclusion] After hydroxyl esterification, the insecticidal activity of mesaconitine was declined and the existence of hydroxyl at 3rd position in mesaconitine played very important influences on insecticidal activity.

A Preliminary Study on Biological Activity of Wikstroemia chamaedaphne Meissn

[ Objective ] The insecticidal and antibacterial bioactivity of Wikstroemia chamaedaphne Meissn were screened and bioactive substances in it were separated and purified. [ Method] The Wikstroemia chamaedaphne Meissn was conducted ultrasonic extraction in petroleum ether, ethyl acetate and methanol. The insecticidal activity of Wikstroemia chamaedaphne Meissn to Mythimna separata walker and aphid were determined. The antibacterial activity of Wikstroemia chamaedaphne Meissn to Fusarium graminearu, Glomerella cingulata, F. oxysporium f. sp niveum, Alternaria solani and Fusarium oxysporium were also determined. The bioactivity-guided methods such as opencolumn chromatography and Pre-HPLC method were used to separate active components in petroleum ether extract from Wikstroemia chamaedaphne Meissn. [ Result] When the concentration was 500 mg/L, 3 kinds of extracts from Wikstroemia chamaedaphne Meissn didn＇ t show obvious antibacterial bioactivity to 5 kinds of test samples. When the concentration was 5%, petroleum ether extract show certain topical toxicity to aphids. The ethyl acetate extract showed certain antifeedant activity to 3^rd instar Larvae of Mythimna separata Walker. The fraction F4 of petroleum ether extract possessed highest topical toxicity to aphids and the lethality was 60.00%. [ Conclusion] Wikstroemia chamaedaphne Meissn contained many insecticidal constituents whose active parts and mechanism were needed further researches.

Expression, Purification and Insecticidal Activity Analysis of Cry1Ac without Helix α-1

[Objective] This study aimed to analyze the expression, purification and insecticidal activity of Cry1Ac without helix α-1 of domain I in E. coli. [Method] Plasmid of B. thuringiensis HD-1 was used as the template for amplification of Cry1Ac gene, which was linked with pET28a vector to construct the recombinant plasmid. After transformation and IPTG induction, expression of target protein was detected by using SDS-PAGE method. Target protein was extracted and coated on the surface of feed for H. armigera to determine the insecticidal activity. [Result] Cry1Acdela1 gene could normally express the target protein in E. coli. However, the target protein existed in the form of inclusion body. Results of bioassay analysis showed that under the same concentration, fatality rate of activated Cry1Ac toxin reached above 75%, while that of Cry1Acdelα1 was only about 10%. [Conclusion] E. coli-expressed Cry1Acdelα1 had no insecticidal activity against H. armigera.

Antifeedant Activity of Three Plant Extracts Including P. acinosa against Pieris rapae

[Objective] The research aimed to study antifeedant activity of Phytolacca acinosa Roxb., Setaria viridis （L.） Beauv and Viola yedoensis Makino extracts against Pieris rapae. [Method] Activity material was extracted from S. viridis （L.）, P. acinosa and V. yedoensis using acetone cold soak method, and non-selective antifeedant activity of extracts to Pieris rapae larva was determined by using lobular plate addition method. [Result] The results showed that the acetone leaching agent of P. acinosa had most obvious antifeedant effects on Pieris rapae. The antifeedant rate were 74.53% and 82.34% at 24 and 48 h respectively. With the concentration increasing, the antifeedant effect of P. acinosa extracts increased. The antifeedant rate of 0.050 g/ml treatment was the highest, being 74.53% and 82.34% at 24 and 48 h. [Conclusion] P. acinosa could be studied and utilized as potential botanical insecticide.

Larvicidal activity and microencapsulation of tobacco(Nicotiana tabacum)extract on Malacosoma neustria testacea larvae

To understand and improve the stability of the insecticidal activity of tobacco extract,the 3 rd instar larvae of Malacosoma neustria testacea was determined by the leaf film method.Spectrophotometry identified extract effects on activities of several enzymes.In addition,to improve the stability of the extract,microcapsules were prepared by complex coacervation and phase separation with the extract as core material,and gelatin and gum arabic as wall material.With the embedding rate as the evaluation index,the response surface method was used to optimize the preparation process of the microcapsules.The results show that the extract had a strong insecticidal activity on the larvae,with inhibitory effects on several enzymes examined of carboxylesterase,acetylcholinesterase,glutathione-S transferase,catalase,and superoxide dismutase.The inhibition rate increased with time.The best preparation process of tobacco extract microcapsules was 25% mass fraction of emulsifier,2.05% mass fraction of gelatin,3% mass fraction of gum arabic,1.34 wall core ratio,36 min of complex coacervation time.The embedding rate was 58.4% which is approximately the theoretical embedding rate(58.9%).The microcapsules prepared by this method have a smooth surface,good combining form and particle size distribution,and a median diameter of 8.6 μm.Infrared characteristic peaks of the extracts were preserved at 877.55 cm^(-1) and2922.13 cm^(-1).Microencapsulation can improve the thermal stability of the tobacco extract.Indoor toxicity tests showed that LC_(50) of extract microcapsules was 20.2 mg·mL^(-1),equivalent to the toxicity level of the tobacco extract itself,indicating that microencapsulation did not reduce extract insecticidal effects.This research may provide a reference for the optimization of the tobacco extract microcapsule preparation process.

Diamide derivatives containing a trifluoromethylpyridine skeleton:Design,synthesis,and insecticidal activity

Diamide derivatives are biologically active molecules that have been widely applied in recent years in research on pesticides,especially insecticides.Using a simple and environmentally friendly scheme,a series of new diamide derivatives containing a trifluoromethylpyridine skeleton was designed,synthesized,and confirmed by^(1)H,^(19)F and^(13)C NMR,and HR-MS.Their insecticidal activities against Plutella xylostella and Helicoverpa armigera were measured and the relationship between structure and activity was investigated.Eight of the title compounds(D2,D5,D10,D21,D28,D29,D30 and D33)showed 100%activity against P.xylostella at 500 mg L^(-1).One compound,D33,still showed 100%activity against P.xylostella at 100 mg L^(-1)and had the lowest LC_(50)(lethal concentration 50%,3.7 mg L^(-1))among the synthesized compounds.Molecular docking analysis revealed that D33 could be thoroughly embedded in the active pocket of the ryanodine receptor via hydrogen bonding in a manner similar to the commercial insecticide chlorantraniliprole.

Purification, characterization and evaluation of insecticidal activity of trypsin inhibitor from Albizia lebbeck seeds

A Bowman-Birk inhibitor with activity against gut proteases of Helicoverpa armigera was extracted in 0.I M sodium phosphate buffer from defatted seed flour of Albizia lebbeck. It was purified to 29.62 folds with 51.43% recovery using ammonium sulfate precipitation, gel filtration chromatography on Sephadex G-100 column and ion ex- change chromatography on DEAE-Sephadex As0. The purified protein had a molecular weight of 12,303 daltons as determined by SDS-PAGE. It was found to be heat stable up to 60~C and had two pH optima of 7.5 and 9.0. The inhibitor exhibited non-competitive pattern of inhibition with a low Ki value of 0.2 ~tM. The inhibitoi- was found to be susceptible to varying concentrations of reducing agents like DTT and 2- mercaptoethanol, thereby indicating the role of disulphide bridges in maintaining its three dimensional structure and stability. The purified inhibitor caused mortality and suppressed larval growth ofPieris brassi- cae larvae. It was also found to be effective against gut trypsin extracted from Spodoptera littoralis. The sequence of the genes encoding for such inhibitors can be determined and the genes expressing protease inhibitors can be used in vegetable crops to confer resistance against insect pests and other plant pathogens.

Synthesis,Crystal Structure and Insecticidal Activity of N-(pyridin-2-ylmethyl)-1-phenyl-1,4,5,6,7,8-hexahydrocyclohepta[c]pyrazole-3-carboxamide

The title compound N-（pyridin-2-ylmethyl）- 1-phenyl- 1,4,5,6,7,8-hexahydrocy-clohepta[c]pyrazole-3-carboxamide 5 （C21I-I22N40, Mr = 346.42） has been synthesized andstructurally characterized by IR, 1H NMR, 13C NMR, H RMS and single-crystal X-raydiffraction. The crystal crystallizes in monoclinic system, space group P21/n with a = 8.668（2）,b = 22.236（4）, c = 9.539（2） A, β = 108.68（3）°, V = 1786.4（7）/k3, Z = 4, Dx= 1.288 g/cm3,F（000） = 736,μ（MoKa） = 0.649 mm^-1, the final R = 0.0354 and wR = 0.0933 with 3234observed reflections with I 〉 2σ（/）. The benzene and pyrazole rings are nearly coplanar with adihedral angle of 50.977（46）°. The dihedral angle between the central pyrazole and pyridinerings is 11.688（46）°. No classical hydrogen bonds were found in the molecules. Two adjacentmolecules in crystal packing of compound 5 were linked by two intramolecularhydrogen-bonding interactions C（15）-H（15）…O（1） to generate a stable structure. Compound 5had weak insecticidal activity against the diamondback moth （Plutella xylostella）, butexhibited good activity against cotton bollworm （Helicoverpa armigera）.

Synthetic derivatives of spiromesifen and their bioactivity research

Sixteen new derivatives of spiromesifen were synthesized from 3-（2,4,6-trimethylphenyl）-4-hydroxy-△^3-dihydrofuran-2-one. Their bioactivities against diamondbackmoth （Plutella xylostella） and spider mites （Tetranychus cinnabarinus） were also evaluated. The structures of these derivatives were confirmed by ^1H NMR, MS.

Study on Chemical Synthesis and Insecticidal Activity of Peptideκ-CTx-btg02

[Objectives]This study was conducted to investigate the chemical synthesis of peptideκ-CTx-btg02.[Methods]Linear peptideκ-CTx-btg02 was synthesized by solid phase peptide synthesis(SPPS).After oxidation and folding of the linear peptide,mass spectrometry identification and high performance liquid chromatography(HPLC)purification were performed.Then,the MTT method and insect injection method were applied to study its insecticidal activity.[Results]The peptideκ-CTx-btg02 was successfully synthesized by the SPPS method,and identified for the formation of disulfide bonds by mass spectrometry identification,and the purity after HPLC separation was greater than 95%.The MTT experiment showed that the peptideκ-CTx-btg02 could inhibit the growth of insect cells sf9,with a half effective dose of 0.13 nM.The insect injection experiment showed that the peptideκ-CTx-btg02 could effectively kill Tenebrio molitor,with a half lethal dose of 15.6 nM.The results of the electrophysiological experiment showed that 10μM peptideκ-CTx-btg02 had no blocking activity on murine acetylcholineα2β3 andα3β4.Therefore,the peptideκ-CTx-btg02 had a good inhibitory effect on the growth of insect cells,highly effective insecticidal activity and weak mammalian toxicity.[Conclusions]This study lays a foundation for the development of new,safe and efficient peptide biological insecticides.

Ethnobotanical Investigation of Insecticidal Plants and Evaluation of the Insecticidal Activity of Capsicum chinense on Sitophilus zeamaïs of Maize and Aphis craccivora of Groundnuts: Case of Boffa, BokéRegion (Republic of Guinea)

The present study, which aims to contribute to the valorization of Capsicum chinense, focuses on ethnobotanical investigations and evaluation of insecticidal activity on Sitophilus zeamaïs and Aphis craccivora, as well as the germination power of infected maize and peanut grains treated and not treated with Capsicum chinense leaf powder. To do this, the moisture content of corn and peanuts was determined by the techniques: drying in the sun and in the oven. The insecticidal effect of the powder was tested on maize and peanut kernels infected by the AGRAR (2013) method, followed by a test of the germination potency of these treated grains. The average moisture values of corn and groundnut kernels are 14.51% and 12.25% respectively. The results show, a higher insecticidal efficacy of Capsicum chinense leaf powder on Sitophilus zeamaïs and Aphis craccivora with doses (2, 4 and 8 g). The average germination rates of maize and peanut kernels infected and treated with this powder are: (16.5%;63%;99.5%) and (21.25%;96.5%;99.75%), respectively. This study could be a bio-insecticide alternative for the peasant community in the conservation of cereal and legume products.

Larvicidal activity of Neem oil and three plant essential oils from Senegal against Chrysodeixis chalcites(Esper, 1789)

Objective: To evaluate the insecticide, larvicidal and repellent activity of the essential oils from Callistemon viminalis, Melaleuca leucadendron, and Hyptis suaveolens against Chrysodeixis chalcites and to compare it with neem oil(Azadirachta indica). Methods: The essential oils of the leaves of these aromatiques plants were extracted by steam distillation and contacts tests were carried out. Results: Essential oils in ethanol from Callistemon viminalis showed a higher biological activity than the neem with 100% larval mortality at the concentration of 2 μg/m L for 6 h, 100% and 90% in ethanol from Melaleuca leucadendron and Hyptis suaveolens, respectively at the concentration of 4 μg/m L for 24 h. By inhalation, the essential oils from Melaleuca leucadendron and of Hyptis suaveolens were more effective with mortality rates of larvae 100% and 50% respectively at 2 μg/L air applied after 24 h. Nevertheless, the neem has shown to be a repulsive plant and anti-nutritional plant. A significant difference in the percentages of consumption between leaves treated with neem oil and the control samples was observed(Newman-Keuls test) except for Melaleuca leucadendron. Conclusions: The results of the study highlight remarkable biocide, properties of tested extracts, which provides important opportunities for the development of biopesticides.

Regioselective Synthesis, Crystal Structure and Biological Activity of Novel 2-Phenylsulfonylhydrazono-3-(3-trifluoromethylphenyl)-2(3H)-thiazoline Derivatives

The title compounds 2-phenylsulfonylhydrazono-3-（3-trifluoromethylphenyl）-2（3H）- thiazoline derivatives were designed and synthesized via the reaction of thiosemicarbazides with chloroacetaldehyde, and their chemical structures were characterized by ^1H NMR, ^13C NMR, elemental analysis and MS. Furthermore, 4a was characterized by single-crystal X-ray diffraction. The target compound 4a （C16H12F3N302S2, Mr = 399.41） crystallized in monoclinic system, P21/c space group with a = 7.7457（4）, b = 13.5850（7）, c = 16.5455（10） A, β= 99.698（2）°, V= 1656.30（16） A3, Dc = 1.602 g/cm^3, Z = 4, F（000） = 816, λ = 0.71070 A,μ（MoKa） = 0.370 mm^-1, R = 0.072 and wR = 0.0867. The crystal structure of 4a revealed the cyclization of thiosemicarbazide moiety with chloroacetaldehyde at the N（l） position. The title compounds exhibited good insecticidal activity against spider mite （Tetranychus cinnabarinus Boisduval） and favorable fungicidal activity against Corynespora cassiicola.

Insecticidal Activity of Crystals and Spores of Purified Bacillus thuringiensis 00-50-5 Strain against Trichoplusia ni Hubner

[ Objective] The paper was to study pathogenic mechanism and action mode of Bacillus thuringiensis （Bt） 00-50-5 strain against Trichoplusia ni Hub- ner. [Method] The crystals and spores of B. thuringiensis 00-50-5 strain were first isolated and purified, and their insecticidal activities against T. ni were com- pared. [ Result] Either the mixture of pure crystals and pure spores of 00-50-5 strain, or only pure crystals could kill T. n/larvae under the concentration of 100 p.g/mL after 48 h, and the mortality rate reached 100% ; pure spores could not kill the larvae. Determination results of median lethal concentration showed that in- secticidal activity of pure crystal of Bt 00-50-5 strain （LGs0 =0.32 p~/mL） was higher than the mixture of crystals and spores （LCs0 =0.48）, but the insecticidal acti^ty of pure spores was very low （LC50 〉 500.00）. Therefore, the crystals were primarily responsible for causing death of larvae. [ Conclusion] The paper provides theoretical basis for application of Bt 00-50-5 strain.

Insecticidal Activity of Piperine Isolated from Piper sarmentosum

In order to clarify the insecticidal active ingredients of Piper sarmentosum,one active ingredient was isolated from ethanol extract of P. sarmentosum by bioassay-guided fractionation method. Its chemical structures were identified to be piperine by MS,1H NMR,13C NMR. The insecticidal activity of piperine and ethanol extract of P. sarmentosum against Aleurodicus dispels Russell were tested by leaf dip method. The results showed that piperine and ethanol extract of P. sarmentosum exhibited strong insecticidal activity against adults and nymphs of A. dispels; LC50values against adults were 28. 59 and 224. 31 mg/L,and LC50values against nymphs were 65. 91 and 336. 68 mg/L,respectively. There was no significant difference between piperine and azadirachtin against adults and nymphs of A.dispels. Therefore,piperine might be one of the main insecticidal ingredients of P. sarmentosum. In addition,piperine and ethanol extract showed ovicidal activity with different mode of action,piperine reduced the survival rate of newly hatched nymphs while ethanol extract impacted hatch of eggs.

Synthesis analogues of milbemycin and their bioactivity evaluation

Eight new 13-O-aminocarbonylivermectin aglycones and 4＇-O-aminocarbonylivermectin monosaccharide were synthesized from ivermectin agiycone and ivermectin monosaccharide by the selective protection of C5-OH group.Their bioactivities were evaluated against spider mites（Tetranychus cinnabarimts）,aphid（Aphis fabae） and oriental armyworm（Mytliimma sepatara）. Their structures were confirmed by ^1H NMR.MS.

Preparation and Insecticidal Activity of Sea Anemone Peptide AP-GI from Aiptasia pallida

[Objectives] This study was conducted to synthesize sea anemone peptide Ap-GI and investigate its insecticidal activity. [Methods] The linear peptide Ap-GI was synthesized by solid phase peptide synthesis(SPPS), and the linear peptide was subjected to the two-step oxidative folding, mass spectrometry identification and high performance liquid chromatography(HPLC) purification. Then, the MTT method and insect injection method were used to study its insecticidal activity. [Results] The synthesized sea anemone peptide had a purity of 95%. The test results of the MTT method showed that the peptide Ap-GI had the activity of inhibiting the growth of insect cells sf9 with the median effective dose of 0.7 nM;and the test results of the injection method on yellow mealworms showed that the peptide Ap-GI had high insecticidal activity, and the median lethal dose was 16.9 nM. [Conclusions] The sea anemone peptide Ap-GI from Aiptasia pallida has a good inhibitory effect on the growth of insect cells and high-efficiency insecticidal activity, which can lay a foundation for the development of new, safe and efficient peptide biological insecticides.

Preparation of Sea Anemone Peptide Toxin Ap-TxI and Its Insecticidal Activity

[Objectives]This study was conducted to synthesize sea anemone peptide toxin Ap-TxI and investigate its insecticidal activity. [Methods] The sea anemone linear peptide toxin Ap-TxI was synthesized by the solid-phase peptide synthesis(SPPS), and six cysteines were oxidized to form three disulfide bonds by a three-step directional oxidation method. Then, purification by high performance liquid chromatography(HPLC) and mass spectrometry identification were performed. Finally, the insect cytotoxicity and insecticidal activity of Ap-TxI were studied by the MTT method and insect injection method. [Results] The oxidized peptide Ap-TxI with three disulfide bonds in natural configuration was successfully synthesized by the SPPS method, and its purity was >90% by HPLC analysis. The results of the MTT method showed that Ap-TxI was active on the growth of insect cells sf9, with a half effective dose of 0.2 nM;and the results of the mealworm injection test showed that the polypeptide Ap-TxI had high insecticidal activity with a median lethal dose of 11.7 nM. [Conclusions] The sea anemone peptide toxin Ap-TxI with high insecticidal effect was obtained, laying a foundation for the development of new, efficient and safe biological insecticides.