Loop-Mediated Isothermal Amplification for Detecting Gly-4891-Glu and Ile-4734 Multiple Mutations of Ryanodine Receptor in the Fall Armyworm, Spodoptera frugiperda.

The key mutations, such as the Gly-4891-Glu substitution and the Ile-4734 multiple substitutions within the ryanodine receptors (RyR), are linked to diamide resistance in fall armyworm (FAW), Spodoptera frugiperda. In this study, we found that FAW remained sensitive to cyantraniliprole and chlorantraniliprole, while its sensitivity to flubendiamide was reduced. Moreover, a low level of heterozygous mutation at I4743 was observed. To facilitate the detection procedure of these mutations, a simple and efficient loop-mediated isothermal amplification (LAMP) protocol was developed for operation. The reaction for detecting the G4891E and I4743 single or multiple mutations was carried out at 68 °C for 85 min and 68 °C for 85 min or 68 °C for 65 min, respectively. These LAMP reactions can be easily observed via visualization of the color change from pink to yellow. This assay provides a simple, convenient, and effective means of detecting mutations in the RyR of FAW for pest management purposes.

Sublethal effect and detoxifying metabolism of metaflumizone and indoxacarb on the fall armyworm, Spodoptera frugiperda.

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera, Noctuidae), is a highly polyphagous invasive pest that damages various crops. Pesticide control is the most common and effective strategy to control FAW. In this study, we evaluated the toxicity of metaflumizone and indoxacarb against third-instar FAW larvae using the insecticide-incorporated artificial diet method under laboratory conditions. Both metaflumizone and indoxacarb exhibited substantial toxicity against FAW, with LC50 values of 2.43 and 14.66 mg/L at 72 h, respectively. The sublethal effects of metaflumizone and indoxacarb on parental and F1 generation FAW were investigated by exposing third-instar larvae to LC10 and LC30 concentrations of these insecticides. Sublethal exposure to these two insecticides significantly shortened adult longevity, extended pupal developmental times and led to reduced pupal weight, pupation rates, and adult fecundity in the treated parental generation and F1 generation at LC10 or LC30 concentrations, in comparison to the control group. The larval developmental times were shortened in the parental generation but prolonged in the F1 generation, after being treated with sublethal concentrations of metaflumizone. Furthermore, larvae exposed to LC10 or LC30 concentrations of indoxacarb exhibited elevated activity levels of cytochrome P450 monooxygenase and glutathione S-transferase, which coincides with the observed synergistic effect of piperonyl butoxide and diethyl maleate. In conclusion, the high toxicity and negative impact of metaflumizone and indoxacarb on FAW provided significant implications for the rational utilization of insecticides against this pest.

Identification and up-regulation of three small heat shock proteins in summer and winter diapause in response to temperature stress in Pieris melete.

Small heat shock proteins (sHSPs) are conserved proteins that play key roles in organismal adaptation to adversity stressors. However, little is known about sHSPs during summer diapause. Three sHSP genes: PmHSP19.5, PmHSP19.9, and PmHSP20.0 were identified and cloned from Pieris melete. Sequence alignment and phylogenetic analysis revealed that the three sHSPs have a typical, conserved α-crystallin domain. PmHSP19.5 and PmHSP20.0 were both upregulated in summer diapause (SD) and winter diapause (WD), compared to non-diapause (ND) pupae. All three sHSPs were upregulated and showed similar trends in response to thermal stress. The 0 °C chilling treatment slightly affected sHSP transcripts in ND pupae, whereas both PmHSP19.5 and PmHSP19.9 were upregulated and PmHSP20.0 was downregulated after chilling at 0 °C for 24-96 h in both SD and WD pupae. The transcripts of PmHSP19.5 and PmHSP19.9 were significantly induced at 31 °C for 30 d in SD and WD pupae. The PmHSP20.0 transcript gradually decreased during the SD and WD programs. This is the first time that sHSPs have been linked to both overwintering and summer diapause processes. These findings suggest that sHSPs are involved in both summer and winter diapause maintenance and play a possible key role in temperature stress.

Effects of starvation on respiratory metabolism and energy metabolism in the cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

Intermittent food shortages are commonly encountered in the wild. To cope with the threat of starvation, insects initiate a suite of behavioral activities and physiological countermeasures. The cotton bollworm, Helicoverpa armigera, is a major agricultural pest worldwide, but how H. armigera modulates its metabolism under starvation remains ambiguous. In the present study, the respiratory rates (V̇O2; ml g-1 h-1) from the third-larval instar to the pupal stage were first determined. Our results highlighted a transient rise during the larval-larval molt and larval-pupal transition, followed by a sharp decline in the pupal stage and, finally, an upward trend before eclosion. When subjected to food deprivation, the starved larvae experienced a significant decline in the rates of O2 consumed and CO2 produced, as well as in respiratory quotient (RQ) values, indicative of severe metabolic depression during starvation and a shift of metabolic substrates with prolonged starvation. For metabolic substrate analysis, an apparent decline in triglyceride and glycogen contents was observed in the starved larvae, and the hemolymph trehalose content was significantly reduced throughout starvation. In addition, comparative transcriptome analysis showed that 48 h of larval starvation caused substantial transcriptional regulations in several energetically costly processes, wherein the marked up-regulations were detected in the pathways of glycolysis and fatty acid metabolism. Overall, our work makes a comprehensive study on the respiratory rate and energy metabolism in the starved H. armigera larvae, and provides a deep insight into the physiological adaptive strategies to alleviate nutritional stress.

[Regulating effect of agricultural landscape pattern on ecological pest control by natural enemies.] / å†œç”°æ™¯è§‚æ ¼å±€å¯¹å®³è™«å¤©æ•Œç”Ÿæ€æŽ§å®³åŠŸèƒ½çš„è°ƒæŽ§ä½œç”¨.

It's well known that regional agricultural landscape pattern can significantly regulate the relationship between plants, herbivorous insects and natural enemies. The improvement of the ecosystem services of regional agricultural landscape pattern has profound impacts on pest control, which has received much interests in current landscape ecology and pest control studies. This review systematically summarized the impacts of agricultural landscape pattern, as well as its succession on the diversity and ecological pests control of important natural enemies. From the perspective of different natural enemies, the impacts of agricultural landscape pattern on natural predators, parasi-toids, beneficial microorganisms and its ecological pests control functions were all recapped. We outlined crop spatial distribution and vegetation habitat management strategies. This review could provide useful insights in improving the ecosystem services of natural enemies in agricultural landscape based on the vegetation habitat management, and offer theoretical support for regional ecological agriculture in response to the strategy of 'chemical pesticide reduction application'.

Molecular characterization of three Hsp90 from Pieris and expression patterns in response to cold and thermal stress in summer and winter diapause of Pieris melete.

Heat shock proteins (Hsps) have been linked to stresses and winter diapause in insects, but whether they are components of summer diapause is still unknown. In this study, complementary DNAs of Hsp90 from Pieris melete, Pieris rapae and Pieris canidia named PmHsp90, PrHsp90 and PcHsp90, respectively, were cloned and sequenced. The deduced amino acid sequence consisted of 718 amino acid residues with a putative molecular mass of 82.6, 82.6 and 82.7 kDa, respectively. The amino acid sequences contained all of the five conserved signature motifs in the Hsp90 family and a bHLH protein folding activity region. The differential expression pattern of PmHsp90 in response to summer diapause and winter diapause, which are related to heat/cold stress, was investigated. Cold stress induced Hsp90 up-regulation in summer and winter diapause pupae, but not in non-diapause individuals. Heat shock up-regulated PmHsp90 gradually with an increase in temperature in summer diapause, and PmHsp90 was rapidly up-regulated in winter diapause. After 30 min heat shock at 39°C, substantial up-regulation of PmHsp90 transcript levels were observed both in summer and winter diapause. However, in non-diapause a relatively stable expression was found under different durations of 39°C heat shock. Compared to the optimal treatment of 18°C for diapause development, a high temperature acclimation of 31°C induced PmHsp90 up-regulation in summer diapause, whereas a low temperature acclimation of 4°C induced up-regulation in winter diapause. The current results indicate that Hsp90 may play an important role in response to heat/cold stress both in summer and winter diapause.