To explore natural product-based pesticide candidates, a series of indole derivatives containing the isoxazoline skeleton at the N-1 position were synthesized by 1,3-dipolar [2 + 3] cycloaddition reaction. Their structures were characterized by melting points (mp), infrared (IR) spectra, proton nuclear magnetic resonance spectra (1H NMR), carbon-13 nuclear magnetic resonance spectra (13C NMR), and high resolution mass spectrometry (HRMS). The single-crystal structures of five compounds were presented. Against Tetranychus cinnabarinus Boisduval, compound 3b showed greater than 3.8-fold acaricidal activity of indole and good control effects under glasshouse conditions. Against Aphis citricola Van der Goot, compounds 3b and 3q exhibited 48.3- and 36.8-fold aphicidal activity of indole and 6-methylindole, respectively. Particularly, compound 3b showed good bioactivities against T. cinnabarinus and A. citricola. Against Eriosoma lanigerum Hausmann, compound 3h and 3i showed 2.1 and 1.9 times higher aphicidal activity compared to indole. Furthermore, the construction of the epidermal cuticle layer of 3b-treated carmine spider mites was distinctly damaged, which ultimately led to their death.

Animals living in clusters should adjust their reproductive strategies to adapt to the social environment. Theories predict that the benefits of cluster living would outweigh the costs of competition. Yet, it is largely unknown how animals optimize their reproductive fitness in response to the changing social environment during their breeding period. We used Tetranychus ludeni Zacher, a haplodiploid spider mite, to investigate how the ovipositing females modified their life-history traits in response to the change of cluster size (i.e., aggregation and dispersal) with a consistent population density (1 ♀/cm2). We demonstrate that (1) after females were shifted from a large cluster (16 ♀♀) to small ones (1 ♀, 5 and 10 ♀♀), they laid fewer and larger eggs with a higher female-biased sex ratio; (2) after females were shifted from small clusters to a large one, they laid fewer and smaller eggs, also with a higher female-biased sex ratio, and (3) increasing egg size significantly increased offspring sex ratio (% daughters), but did not increase immature survival. The results suggest that (1) females fertilize more larger eggs laid in a small population but lower the fertilization threshold and fertilize smaller eggs in a larger population, and (2) the reproductive adjustments in terms of egg number and size may contribute more to minimize the mate competition among sons but not to increase the number of inhabitants in the next generation. The current study provides evidence that spider mites can manipulate their reproductive output and adjust offspring sex ratio in response to dynamic social environments.

To discover new potential pesticide candidates, recently, structural modification of natural bioactive products has received much attention. In this work, a series of new piperine-type ester derivatives were regio- and stereoselectively synthesized based on a natural alkaloid piperine isolated from Piper nigrum. Their structures were characterized by IR, mp, 1H NMR (13C NMR), and high-resolution mass spectrometry (HRMS). Against Tetranychus cinnabarinus Boisduval (Acari: Tetranychidae), compounds 4e, 4f, 4u, and 4v displayed the most significant acaricidal activity with LC50 values of 0.155, 0.117, 0.177, and 0.164 mg/mL, respectively. Particularly, compound 4f showed >120-fold higher acaricidal activity than piperine (LC50: 14.198 mg/mL). Notably, the acaricidal activity of 4f was equivalent to that of the commercial acaricide spirodiclofen (LC50: 0.115 mg/mL). Additionally, against Eriosoma lanigerum Hausmann (Hemiptera: Aphididae), compounds 4w and 4b\' showed 1.8-fold aphicidal activity of piperine. Furthermore, via the scanning electron microscope (SEM) imaging method, the obvious destruction of the construction of the cuticle layer of 4f-treated T. cinnabarinus was observed. Compound 4f could be further studied as a lead acaricidal agent.

The γ-aminobutyric acid receptor (GABAR) has been identified as a target site of some commonly used insecticides. Our pervious study documented an interesting phenomenon, i.e. GABA accumulation was involved in abamectin resistance in the carmine spider mite, Tetranychus cinnabarinus. However, the mechanism of this phenomenon remains to be clarified. In this study, we investigated the interaction between abamectin and GABAR. Firstly, an artificial increase of GABA content was conducted in T. cinnabarinus and toxicity assays showed that GABA accumulation could indeed increase the tolerance of T. cinnabarinus to abamectin in vivo. Subsequently a GABAR of T. cinnabarinus, RDL2, was expressed in Xenopus oocytes and its sensitiveness to abamectin was detected. The results revealed that RDL2 showed significant responses to a series of GABA concentrations and GABA inhibited the effect of abamectin in vitro, providing direct evidence of the abamectin resistance mediated by GABA content. Our data confirmed that GABAR is the action target of abamectin and the GABA accumulation is one of the mechanisms of abamectin resistance in spider mites.

In continuation of our program to develop natural-product-based pesticidal candidates, matrinic/oxymatrinic amides were obtained through structural optimization of matrine. N\'-(4-Fluoro)phenyl-N-(4-bromo)phenylsulfonyloxymatrinic amide (IIm) showed potent insecticidal activity against Mythimna separata. N-(Un)substituted phenylsulfonylmatrinic acids (3a-c) exhibited promising acaricidal activity against Tetranychus cinnabarinus. By qRT-PCR analysis of nAChR subunits and AChE genes and determination of AChE activity of (un)treated T. cinnabarinus, it suggested that the open lactam ring of matrine and carboxyl group and (4-methyl)phenylsulfonyl of N-(4-methyl)phenylsulfonylmatrinic acid (3b) were necessary for action with α2, α4, α5, and β3 nAChR subunits; compound 3b was an inhibitor of AChE in T. cinnabarinus, and AChE was one possible target of action in T. cinnabarinus against 3b; and compound 3b may be an antagonist of nAChR and AChE in T. cinnabarinus.

This is the first systemic assessment of the stereoselectivity of etoxazole enantiomers. Etoxazole\'s stereoselective bioactivity was assessed against target organisms ( Tetranychus urticae eggs and Tetranychus cinnabarinus eggs), and its acute toxicity was assessed toward nontarget aquatic organisms ( Daphnia magna and Danio rerio). Additionally, stereoselective elimination was investigated in three species of fruits (grape and strawberry grown in a greenhouse and apple grown in an open field) and in field soil. The ovicidal activity of (+)-( S)-etoxazole against Tetranychus urticae and Tetranychus cinnabarinus eggs was about 16 and 24 times higher, respectively, than that of (-)-( R)-etoxazole. Inconsistent order of etoxazole isomer toxicity was found toward different aquatic organisms: (+)-( S)-etoxazole showed nearly 8.7 times higher acute toxicity than (-)-( R)-etoxazole toward Daphnia magna, whereas (-)-( R)-etoxazole was ∼4.5 times more toxic to Danio rerio than (+)-( S)-etoxazole. Stereoselective degradation of etoxazole enantiomers showed significant variation in various fruits and field soil. The (+)-( S)-etoxazole was preferentially dissipated in grape and strawberry fruits grown under greenhouse condition, whereas (-)-( R)-etoxazole degraded faster than its antipode in apple fruits and soils under open-field condition. Overall, the stereoselectivity of etoxazole enantiomers should be fully considered in comprehensive environmental health risk in future work.

Thirty phenolic ether derivatives of scopoletin modified at the 7-hydroxy position were synthesized, and their structures were confirmed by IR, ¹H-NMR, 13C-NMR, MS and elemental analysis. Preliminary acaricidal activities of these compounds against female adults of Tetranychus cinnabarinus (Boisduval) were evaluated using the slide-dip method. The results indicated that some of these compounds exhibit more pronounced acaricidal activity than scopoletin, especially compounds 32, 20, 28, 27 and 8 which exhibited about 8.41-, 7.32-, 7.23-, 6.76-, and 6.65-fold higher acaricidal potency. Compound 32 possessed the the most promising acaricidal activity and exhibited about 1.45-fold higher acaricidal potency against T. cinnabarinus than propargite. Statistically significant 2D-QSAR model supports the observed acaricidal activities and reveals that polarizability (HATS5p) was the most important parameter controlling bioactivity. 3D-QSAR (CoMFA: q² = 0.802, r² = 0.993; CoMSIA: q² = 0.735, r² = 0.965) results show that bulky substituents at R₄, R₁, R₂ and R₅ (C₆, C₃, C₄, and C₇) positions, electron positive groups at R₅ (C₇) position, hydrophobic groups at R₁ (C₃) and R₂ (C₄), H-bond donors groups at R₁ (C₃) and R₄ (C₆) will increase their acaricidal activity, which provide a good insight into the molecular features relevant to the acaricidal activity for further designing novel acaricidal agents. Molecular docking demonstrates that these selected derivatives display different bide modes with TcPMCA1 from lead compound and they interact with more key amino acid residues than scopoletin. In silico ADME properties of scopoletin and its phenolic ether derivatives were also analyzed and showed potential to develop as good acaricidal candidates.

In continuation of our program aimed at the development of natural product-based pesticidal agents, three series of novel camptothecin derivatives were designed, synthesized, and evaluated for their biological activities against T. Cinnabarinus, B. brassicae, and B. xylophilus. All of the derivatives showed good-to-excellent activity against three insect species tested, with LC50 values ranging from 0.00761 to 0.35496 mmol/L. Remarkably, all of the compounds were more potent than CPT against T. Cinnabarinus, and compounds 4d and 4c displayed superior activity (LC50 0.00761 mmol/L and 0.00942 mmol/L, respectively) compared with CPT (LC50 0.19719 mmol/L) against T. Cinnabarinus. Based on the observed bioactivities, preliminary structure-activity relationship (SAR) correlations were also discussed. Furthermore, a three-dimensional quantitative structure-activity relationship (3D-QSAR) model using comparative molecular field analysis (CoMFA) was built. The model gave statistically significant results with the cross-validated q2 values of 0.580 and correlation coefficient r2 of 0.991 and  of 0.993. The QSAR analysis indicated that the size of the substituents play an important in the activity of 7-modified camptothecin derivatives. These findings will pave the way for further design, structural optimization, and development of camptothecin-derived compounds as pesticidal agents.

Adapting to specific hosts often involves trade-offs that limit performance on other hosts. These constraints may either lead to narrow host ranges (i.e. specialists, able to exploit only one host type) or wide host ranges often leading to lower performance on each host (i.e. generalists). Here, we combined laboratory experiments on field populations with experimental evolution to investigate the impact of adaptation to the host on host range evolution and associated performance over this range. We used the two-spotted spider mite, Tetranychus urticae, a model organism for studies on the evolution of specialization. Field mite populations were sampled on three host plant species: tomato, citrus tree and rosebay (Nerium oleander). Testing these populations in the laboratory revealed that tomato populations of mites could exploit tomato only, citrus populations could exploit citrus and tomato whereas Nerium populations could exploit all three hosts. Besides, the wider niche ranges of citrus and Nerium populations came at the cost of low performance on their non-native hosts. Experimental lines selected to live on the same three host species exhibited similar patterns of host range and relative performance. This result suggests that adaptation to a new host species may lead to wider host ranges but at the expense of decreased performance on other hosts. We conclude that experimental evolution may reliably inform on evolution in the field.

By using slide-dip bioassay method under laboratory condition, an investigation was made on the lethal effects of scopoletin, bisdemethoxycurcumin (BDMC), and their combination at an optimal mass ratio (7:6) of scopoletin to BDMC against the adult females of Tetranychus cinnabarinus Boisd. A time-dose mortality model (TDM) was established, which passed the Hosmer-Lemeshow test. The sensitivity of the adult females to the concentration change of the acaricides was in the sequence of scopoletin > optimal mass ratio of scopoletin to BDMC > BDMC. The peak mortality of the female adults was found at 32, 28 and 32 h after treated with BDMC, scopoletin, and their combination at the optimal mass ratio, respectively. The values of the LC50 and LC90 at 48 h after treated with BDMC, scopoletin, and their combination at the optimal mass ratio were 0.3324, 0.2035 and 0.2195 mg x mL(-1), and 2.1198, 0.9521 and 1.1617 mg x mL(-1), and the median lethal time (LT50) of BDMC, scopoletin, and their combination at the optimal mass ratio was 7.4, 6.0 and 6.1 h at the concentration 1.0 mg x mL(-1), and 6.4, 4.8 and 5.0 h at the concentration 2.0 mg x mL(-1), respectively. The acaricidal activity and time-dose response of the optimal combination of scopoletin and BDMC were closer to those of scopoletin, suggesting a synergistic acaricidal activity of the combination of scopoletin and BDMC, which was worthy to be developed for application.