Cloning,characterisation and expression profiling of the cDNA encoding the ryanodine receptor in diamondback moth,Plutella xylostella (L.) (Lepidoptera: Plutellidae)

BACKGROUND: The rynodine receptors (RyRs) are the main targets of diamide insecticides such as chlorantraniliprole. To provide the basis for a good understanding of the molecular mechanisms of diamide insecticide resistance, an RyR gene from Plutella xylostella was cloned and characterised in the present paper. RESULTS: A full‐length cDNA sequence of RyR was cloned from P. xylostella through RT‐PCR and rapid amplification of cDNA ends (RACE). The gene (named PxRyR1) is 15 753 bp long, with an open reading frame of 15 354 bp, encoding a predicted RyR of 5117 amino acids. An alternative splicing of the PxRyR1 was also cloned and named PxRyR2. The PxRyR1 shares 77–93% identity with other insect RyRs. Quantitative real‐time PCR analysis showed that the PxRyR was expressed at a high level in second‐instar larvae and adults, at a low level in prepupae and pupae and abundantly in the body wall muscle and head (respectively 6.00 and 3.12 times the expression in the gut). Western blot analysis with anti‐RyR antibodies showed that the RyR was mainly present in the body wall muscle and head, but barely present in the haemocyte and gut. CONCLUSIONS: There are at least two alternative splices of PxRyR expressed in all developmental stages and tissues in P. xylostella at various levels. The results provided the basis for further understanding of the mechanisms of resistance to diamide insecticides in P. xylostella. Copyright © 2012 Society of Chemical Industry     

Design, synthesis and insecticidal activities of novel pyrazole amides containing hydrazone substructures

BACKGROUND: Pyrazole and hydrazone derivatives possess good insecticidal activities; their substructural units are widely used in pesticide design. In an effort to discover new molecules with good insecticidal activities, a series of pyrazole amide derivatives containing hydrazone substructures were synthesised and bioassayed. RESULTS: Bioassays demonstrated that some of the title compounds exhibited notable control of Plutella xylostella (Linnaeus), Helicoverpa armigera (Hübner), Culex pipiens pallens, Laphygma exigua (Hübner), Spodoptera litura (Fabricius), Nilaparvata lugens (Stål) and Rhopalosiphum maidis (Fitch) at 5, 10, 0.25, 200, 20, 100 and 500 mg L^?1 respectively. Comparative molecular field analysis (CoMFA) based on the bioactivities for P. xylostella was studied; the values of q² and r² for the CoMFA model were 0.701 and 0.964 respectively. CONCLUSION: Some of the title compounds displayed good and broad‐spectrum insecticidal activities against different insect species; the CoMFA model revealed that a bulky and negatively charged group at the 4‐position of benzene could enhance insecticidal activity. These results could provide useful information for the design of novel insecticide containing substructural units of pyrazole amide and hydrazone. Copyright © 2011 Society of Chemical Industry     

2-arylalkanoates,a new group of synthetic pyrethroid esters not containing cyclopropanecarboxylates

Although structure modifications of natural pyrethrin constituents have disclosed a variety of potent synthetic analogues, all known examples are cyclopropanecarboxylate esters, a grouping that appeared to be essential for insecticidal activity. Some new substituted 2-phenylalkanoates, whose biological activities are of a similar nature and potency to those of conventional pyrethroids, are now reported. 5-Benzyl-3-furylmethyl and 3-phenoxybenzyl 3-methyl-2-phenylbutyrates and their analogues are potent insecticides. Activity is increased on the introduction of appropriate groups into the 3 and/or 4-positions of the aryl ring and the (S)-2-phenylalkanoates are far more active than their (R)-enantiomorphs. Structure/activity relationships are compared with those for conventional pyrethroids. Some of the new series compare favourably with typical insecticides in tests against Musca domestica, Spodoptera litura and Plutella xylostella.     

Anthranilic diamides: A new class of insecticides with a novel mode of action, ryanodine receptor activation

Development of insecticides with unique modes of action is necessary to combat widespread insecticide resistance. A new class of insecticides has been discovered, the anthranilic diamides, that provides exceptional control through action on a novel target, the ryanodine receptor. Anthranilic diamides potently activate this receptor, releasing stored calcium from the sarcoendoplasmic reticulum causing impaired regulation of muscle contraction. Expression of a recombinant Drosophila ryanodine receptor in a lepidopteran cell line confers sensitivity to anthranilic diamides similar to that observed with native receptors. Ligand-binding studies with radiolabeled ryanodine and radiolabeled anthranilic diamide in Periplaneta americana reveal a single, saturable binding site for this chemistry distinct from that of ryanodine. Further, calcium mobilization studies using mammalian cell lines indicate anthranilic diamides exhibit >500-fold differential selectivity toward insect, over mammalian, receptors. Consequently, anthranilic diamides offer a novel pharmacological tool for calcium signaling research in addition to a unique alternative to existing pest-management strategies.     

Synthesis and herbicidal activities of novel 3-(substituted benzyloxy or phenoxy)-6-methyl-4-(3-trifluoromethylphenyl)pyridazine derivatives

BACKGROUND: 4‐(3‐Trifluoromethylphenyl)pyridazine represents a new series of compounds with bleaching and herbicidal activities. RESULTS: A total of 43 novel 3‐(substituted benzyloxy or phenoxy)‐6‐methyl‐4‐(3‐trifluoromethylphenyl)pyridazine derivatives were synthesised, and their bleaching and herbicidal activities were evaluated through Spirodela polyrrhiza and greenhouse tests. Some compounds exhibited excellent herbicidal activities, even at a dose of 7.5 g ha^?1. CONCLUSION: The results showed that a substituted phenoxy group at the 3‐position of the pyridazine ring and the electron‐withdrawing group at the para ‐position on the benzene ring were essential for high herbicidal activity. Copyright © 2011 Society of Chemical Industry     

Cross-resistance patterns and fitness in fufenozide-resistant diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

BACKGROUND: Fufenozide is a novel non‐steroidal ecdysone agonist with good efficacy against diamondback moth (DBM), Plutella xylostella (Lepidoptera: Plutellidae). At present, it is widely applied for the control of a range of lepidopterous pests in China. This study compared the activities of fufenozide and 12 other insecticides against unselected and fufenozide‐selected strains of DBM to examine potential patterns of cross‐resistance. The relative fitness of the fufenozide‐selected strain was assessed to provide information pertinent to insecticide resistance management. RESULTS: Compared with the susceptible strain (JSS), the fufenozide‐resistant strain (JSR) showed high cross‐resistance to dibenzoylhydrazines and benzoylphenylureas, low cross‐resistance to abamectin and no cross‐resistance to organophosphates, carbamates and pyrethroids. JSR had a lower reproductive ability and a relative fitness of 0.5 compared with JSS. CONCLUSION: P. xylostella has the potential to develop resistance to fufenozide, albeit at the expense of fitness. Cross‐resistance between the same and other classes of insecticides is of concern, and should be a key consideration when implementing fufenozide‐based control strategies for this species. Copyright © 2011 Society of Chemical Industry     

Design, synthesis and structure-activity relationship of novel coumarin derivatives

BACKGROUND: The lead coumarin derivative (E)‐methyl 3‐methoxy‐2‐[2‐(4‐methylcoumarin‐7‐yloxymethyl)phenyl]acrylate was discovered by using an intermediate derivatisation method. To discover new coumarin derivatives with improved activity, a series of substituted coumarins were synthesised and bioassayed. RESULTS: The compounds were identified by ¹H NMR, IR, MS and elemental analysis. Bioassays demonstrated that some of the title compounds exhibited excellent fungicidal activity against cucumber downy mildew at 25 mg L^?1. The relationship between structure and fungicidal activity is reported. CONCLUSION: The present work demonstrates that coumarin derivatives containing methoxyacrylate moieties can be used as possible lead compounds for developing novel fungicides. Copyright © 2011 Society of Chemical Industry     

含环丙甲酸类活性片段的新型二酰胺衍生物的设计、合成及生物活性

为探索具有新颖结构的杀虫剂先导,将多种含环丙甲酸的活性片段引入二酰胺类杀虫剂,设计并合成了21个新型的二酰胺类衍生物,其结构均经过核磁共振波谱(NMR)及高分辨质谱(HRMS)表征.测试了所有目标衍生物对苜蓿蚜Aphis craccivora、东方黏虫Mythimna separate、小麦赤霉病菌Fusarium g...     

Residual toxicity and sublethal effects of chlorantraniliprole on Plutella xylostella (lepidoptera: plutellidae)

BACKGROUND: The diamondback moth (DBM), Plutella xylostella (L.), is the most important pest of cruciferous vegetables in the world. Chlorantraniliprole is a novel anthranilic diamide insecticide registered for the control of lepidopteran pests. The dose response, residual toxicity and sublethal effects of chlorantraniliprole applied for 48 h at LC₁₀ (0.02 mg L^?1) and LC₂₅ (0.06 mg L^?1) on P. xylostella were investigated. RESULTS: Leaf‐dip bioassays showed that chlorantraniliprole had a high level of toxicity against larvae of P. xylostella, and the 48 h LC₅₀ values were 0.23 and 0.25 mg L^?1 for a susceptible and field strain respectively. Chlorantraniliprole also had a long‐lasting effect when the larvae were exposed to chlorantraniliprole field sprayed on radish seedlings. Sublethal effects of chlorantraniliprole were indicated by reduced pupation, pupal weight and adult emergence rates. There was also an increase in the duration of female preoviposition period, decreased fecundity and egg hatch and decreased survival rates of the offspring. The mean values of the net reproductive rate (R₀), intrinsic rate of increase (r_m), finite rate of increase (λ) were significantly lower in the treatment than in control groups. CONCLUSIONS: These results indicate that chlorantraniliprole is effective against P. xylostella. The sublethal concentrations of chlorantraniliprole may reduce the population growth of P. xylostella by decreasing its survival and reproduction, and by delaying its development. Copyright © 2012 Society of Chemical Industry     

Quantitative structure-activity relationships of monoterpenoid binding activities to the housefly GABA receptor

BACKGROUND: Monoterpenoids are a large group of plant secondary metabolites. Many of these naturally occurring compounds have shown good insecticidal potency on pest insects. Previous studies in this laboratory have indicated that some monoterpenoids have positive modulatory effects on insect GABA receptors. In this study, the key properties of monoterpenoids involved in monoterpenoid binding activity at the housefly GABA receptor were determined by developing quantitative structure‐activity relationship (QSAR) models, and the relationship between the toxicities of these monoterpenoids and their GABA receptor binding activities was evaluated. RESULTS: Two QSAR models were determined for nine monoterpenoids showing significant effects on [³H]‐TBOB binding and for nine p‐menthane analogs with at least one oxygen atom attached to the ring. The Mulliken charges on certain carbon atoms, the log P value and the total energy showed significant relationships with binding activities to the housefly GABA receptor in these two QSAR models. CONCLUSIONS: From the QSAR models, some chemical and structural parameters, including the electronic properties, hydrophobicity and stability of monoterpenoid molecules, were suggested to be strongly involved in binding activities to the housefly GABA receptor. These findings will help to understand the mode of action of these natural insecticides, and provide guidance to predict more monoterpenoid insecticides. Copyright © 2012 Society of Chemical Industry     

Synthesis of 2-amino-6-oxocyclohexenylsulfonamides and their activity against Botrytis cinerea

BACKGROUND: With the objective of exploring the fungicidal activity of 2‐oxocyclohexylsulfonamides (2), a series of novel 2‐amino‐6‐oxocyclohexenylsulfonamides (6 to 23) were synthesised, and their fungicidal activities against Botrytis cinerea Pers. were evaluated in vitro and in vivo. RESULTS: The compounds were characterised by IR, ¹H NMR and elemental analysis. Bioassay results of mycelial growth showed that compounds 6 to 23 had a moderate antifungal activity against B. cinerea. N‐(2‐methylphenyl)‐2‐(2‐methylphenylamino)‐4,4‐dimethyl‐6‐oxocyclohexenylsulfonamide (13) and N‐(2‐chlorophenyl)‐2‐(2‐chlorophenylamino)‐6‐oxocyclohexenylsulfonamide (21) showed best antifungal activities, with EC₅₀ values of 8.05 and 10.56 µg mL^?1 respectively. Commercial fungicide procymidone provided an EC₅₀ value of 0.63 µg mL^?1. The conidial germination assay showed that most of compounds 6 to 23 possessed excellent inhibition of spore germination and germ‐tube elongation of conidia of B. cinerea. For in vivo control of B. cinerea colonising cucumber leaves, the compound N‐cyclohexyl‐2‐(cyclohexylamino)‐4,4‐dimethyl‐6‐oxocyclohexenylsulfonamide (19) showed a better control effect than the commercial fungicide procymidone. CONCLUSION: The present work demonstrated that 2‐amino‐6‐oxocyclohexenylsulfonamides can be used as possible new lead compounds for further developing novel fungicides against B. cinerea. Copyright © 2011 Society of Chemical Industry     

草地贪夜蛾鱼尼丁受体与双酰胺类杀虫剂的结合模式及抗性风险分析

为研究草地贪夜蛾Spodoptera frugiperda (以下简称FAW) 对双酰胺类杀虫剂的抗性机制,根据FAW鱼尼丁受体 (ranodine receptor,RyR) 跨膜区域的氨基酸序列,通过同源建模的方法构建其三维结构;通过对比FAW与小菜蛾Plutella xylostella (以下简称DBM)RyR跨膜区域的氨基酸序列,发现两者具有较高的同源度,进而根据已报道的FAWRyR的突变位点I4734M及DBMRyR的突变位点G4946E (对应FAWRyR的G4891),在FAWRyR中引入突变,采用分子对接的方法分别分析其突变前后与5种双酰胺类杀虫剂的结合模式。结果表明:FAWRyR的I4734M突变模型与5种双酰胺类杀虫剂的亲和力与野生型的无明显差别,均存在氢键、卤键、疏水作用、π-π相互作用及π-阳离子作用等非共价相互作用;但G4891E突变模型与5种双酰胺类杀虫剂的亲和力下降、形成的作用力方式减少,从而影响RyR与双酰胺类杀虫剂的结合稳定性。本研究分析发现,FAWRyR的G4891E突变可能导致其对双酰胺类杀虫剂产生抗性,需在今后的研究中关注该突变。该研究结果还可会为基于鱼尼丁受体设计杀虫剂提供参考。     

含酰腙结构的新型吡唑酰胺衍生物对小菜蛾的生物活性

采用浸虫法、夹毒叶片法和叶碟法分别测定了13个含酰腙结构的新型吡唑酰胺衍生物对小菜蛾3龄幼虫的触杀、胃毒和拒食活性。结果表明:该类化合物对小菜蛾3龄幼虫具有较高的胃毒和拒食活性,其中苯环上含有氯原子、且酰腙一端的取代基含有杂原子且体积较小的化合物 H7的胃毒和拒食活性最好,明显高于对照药剂毒死蜱。H7 72 h胃毒作用LC50值为0.6 mg/L(毒死蜱的LC50值为7.4 mg/L);有10个化合物的拒食活性高于毒死蜱,其中 H7 48 h的拒食中浓度(AFC50) 最低,为0.6 mg/L,明显低于毒死蜱(AFC50=6.5 mg/L)。供试化合物对小菜蛾3龄幼虫均无触杀活性。     

Molecular cloning and mRNA expression of a ryanodine receptor gene in the cotton bollworm, Helicoverpa armigera

Ryanodine receptors (RyRs) are the targets of novel diamide insecticides. The cotton bollworm, Helicoverpa armigera, is one of the most important cotton pests in the world. In this study, we report the full-length RyR cDNA sequence (named as HaRyR) of H. armigera. The 16,083-bp contiguous sequence encoded 5, 142 amino acid residues, which shares 80% and 78% overall identities with its homologues in Nilaparvata lugens (NlRyR) and Drosophila melanogaster (DmRyR), respectively. All hallmarks of RyR proteins are conserved in the HaRyR, including the GXRXGGGXGD motif conserved in the Ca²⁺ release channels and four copies of RyR domain unique to RyR channels. The previously identified seven lepidopteran-specific RyR residues were also found in HaRyR (N⁴⁹⁷⁷, N⁴⁹⁷⁹, N⁴⁹⁹⁰, L⁵⁰⁰⁵, L⁵⁰³⁶, N⁵⁰⁶⁸ and T⁵¹¹⁹). An amino acid sequence alignment showed that the N-terminal region of HaRyR (residues 188–295) shared high sequence identity with NlRyR (94%) and DmRyR (92%), and moderate sequence identity (47–50%) with three rabbit RyR isoforms, while the short segment of the C-terminal transmembrane region of HaRyR (residues 4632–4676) exhibited moderate sequence identity with NlRyR (69%) and DmRyR (67%), and low sequence identity (19–28%) with three rabbit RyR isoforms. In addition, expression analysis of HaRyR revealed that the mRNA expression level in eggs was significantly lower than in third instar larvae, pupae and adults, and anatomical regulation of HaRyR expression was also observed with the highest expression level in head compared with thorax and abdomen. Our results lay a foundation for comprehensive structural and functional characterization of HaRyR and for understanding of the molecular mechanisms of toxicity selectivity of diamide insecticides among different species.     

Bioassay‐guided isolation of potent aphicidal Erythrina alkaloids against Aphis gossypii from the seed of Erythrina crista‐galli L

BACKGROUND

The cotton aphid (Aphis gossypii Glover) is one of the most invasive pests of cotton. Many botanical phytochemicals have a long history as a source of insecticides, and as templates for new insecticides. This study was undertaken to isolate aphicidal compounds from the seeds of Erythrina crista‐galli L. using the bioassay‐guided isolation method.

RESULTS

Three novel and 11 known Erythrina alkaloids were isolated. Erysodine ( 9 ), erysovine ( 10 ), erysotrine ( 8 ) and erythraline ( 11 ) showed moderate to excellent aphicidal activity with LD₅₀ values of 7.48, 6.68, 5.13 and 4.67 ng aphid^–1, respectively. The Potter spray tower bioassay gave corresponding LC₅₀ values of 186.81, 165.35, 163.74 and 112.78 µg ml^–1. A unique substructure, which presents an sp³ methylene at C‐8, a non‐oxygenated site at N‐9 and a conjugated dienes group (Δ^1,2 and Δ^6,7), plays a crucial role in the aphicidal activity. Application of erythraline ( 11 ) led to different increases in the activities of superoxide dismutase, catalase and glutathione S‐transferase.

CONCLUSION

The study demonstrated that the Erythrina alkaloids erysodine ( 9 ), erysovine ( 10 ), erysotrine ( 8 ) and erythraline ( 11 ) have potential use as botanical aphicides for commercial application, or as templates for the development of new insecticides. © 2017 Society of Chemical Industry     

The metabolism of fenvalerate in plants: The conjugation of the acid moiety

The metabolism of the pyrethroid insecticide fenvalerate [(RS)-α-cyano-3-phenoxybenzyl (RS)-2-(4-chlorophenyl)-3-methylbutyrate] ( I ), and of its most insecticidal (αS,2S) isomer ( II ), has been examined in cabbage plants grown and treated under laboratory conditions with [¹⁴C]chlorophenyl- and [ring-¹⁴C]benzyllabelled preparations of the two compounds. Both insecticides disappeared from the treated leaves with similar half-lives of approximately 12–14 days; they underwent ester cleavage to a significant extent, together with some hydroxylation at the 2- or 4-position of the phenoxy ring, and hydrolysis of the nitrile group to amide and carboxyl groups. Most of the carboxylic acids and phenols thus produced occurred as glycoside conjugates. In separate experiments, the uptake and metabolism of 2-(4-chlorophenyl)-3-methylbutyric acid ( X ), the acidic half of the molecule, were examined in the laboratory, using abscised leaves of kidney bean, cabbage, cotton, cucumber and tomato plants. The acid X was found to be readily converted, mainly into glucose and 6-O-malonylglucose esters in kidney bean, cabbage and cucumber plants, into glucosylxylose, sophorose and gentiobiose esters in cotton, and into two types of triglucose esters with differing isomerism in tomato. One of the acetyl derivatives of the trisaccharide conjugates was identical with the synthetic deca-acetyl derivative of the [1 → 6]-triglucose ester.     

Toxicity and horizontal transfer of chlorantraniliprole against the Asian subterranean termite Coptotermes gestroi (Wasmann): effects of donor:recipient ratio, exposure duration and soil type

BACKGROUND: The effectiveness of chlorantraniliprole and other insecticides (bifenthrin, fipronil, indoxacarb, imidacloprid and chlorfenapyr) were tested against Coptotermes gestroi (Wasmann). Four experiments were conducted: a topical bioassay, a horizontal transfer study, an insecticide bioavailability test and a feeding bioassay. RESULTS: The topical bioassay showed that chlorantraniliprole was significantly less active to C. gestroi at 24 h post‐treatment compared with the other insecticides tested. Nevertheless, it is likely that a lesser amount of chlorantraniliprole was required to cause 50% mortality of C. gestroi at 7 and 14 days post‐treatment. The exposure duration and donor:recipient ratio affect the mortality of recipient termites. Mortality after exposure to chlorantraniliprole in sandy clay was significantly lower than in sand; however, by 14 days, > 90% of donor and recipient termites died in both substrates, irrespective of concentration. Fipronil and imidacloprid showed faster action, and high to moderate toxicity to C. gestroi. Termite workers also ceased to feed after exposure for 1 h to 50 mg kg^?1 chlorantraniliprole‐treated sandy clay. CONCLUSION: Chlorantraniliprole demonstrated delayed toxicity at the lowest label rate (50 mg kg^?1) in sandy clay. Its slow action will enable greater transfer of toxicant between nestmates, while feeding cessation will promote greater social interaction between healthy and exposed termites. Copyright © 2011 Society of Chemical Industry     

Synthesis and insecticidal activities of pyridine ring derivatives of podophyllotoxin

In an attempt to find the biorational pesticides, we synthesized 12 pyridinyl derivatives of podophyllotoxin (PPT) and 4′-demthylepipodophylltoxin (4′-DMEP) in this study. Their structures and the α/β substitution at C-4 were confirmed by ¹H NMR, IR, MS spectral analyses and elemental analysis. The insecticidal activities were tested against fifth-instar larvae of Pieris rapae and the third-instar larvae of Cullex pipiens pallens at concentrations of 250 and 10 μg ml⁻¹. Four derivatives of PPT, 4.1, 4.2, 4.3 and 4.5, showed higher insecticidal activities against P. rapae than PPT, while three derivatives of PPT, 4.4, 4.5 and 4.6, displayed higher mosquito larvicidal activity than PPT, with LC₅₀ values of 1.66, 3.96 and 1.54 mg l⁻¹, respectively. Interestingly, we also found that the pyridine ring derivatives of PPT showed delayed insecticidal activity, which is different from traditional neurotic insecticides. The results suggest that 4′-OCH₃ in the PPT derivatives is essential to keep the insecticidal activity and the insecticidal activities of pyridine ring derivatives of PPT are higher than that of the derivatives of 4′-DMEP, supporting PPT has the potential to be a lead structure of semi-synthetic insecticides.     

Insecticide selectivity to two species of Trichogramma in three different hosts, as determined by IOBC/WPRS methodology

BACKGROUND: The use of Trichogramma species is a potential key strategy in integrated pest management. However, its effectiveness depends on the use of chemicals that do not interfere with parasitism and parasite population growth. Here, a study was made of the effects of synthetic insecticides on Trichogramma pretiosum and Trichogramma exiguum in different hosts (Ephestia kuehniella, Plutella xylostella and Spodoptera frugiperda) and the influence of International Organisation for Biological Control (IOBC/WPRS) methodology in selectivity studies using different Trichogramma species. The insecticides used were commercial formulations (triflumuron at a concentration of 0.2 mL L^?1 water, etofenprox at a concentration of 0.47 mL L^?1 water and endosulfan at a concentration of 7.5 mL L^?1 water); the control treatment consisted of distilled water. Eggs attached to cardboard cards were offered to parasitoids inside glass cages. Parasitised eggs, parasitism and adult emergence rates and parasitism reduction were evaluated. RESULTS: Endosulfan and etofenprox, classified as class‐4 toxic products, were extremely toxic to the parasitoids. Triflumuron, classified as a non‐toxic product, was selective to the parasitoids in eggs of all hosts. CONCLUSIONS: The methodology recommended by IOBC/WPRS influenced results regarding the use of different species of parasitoids, and the use of a single parasitoid species in their experiment is questionable. Copyright © 2011 Society of Chemical Industry