Toxicity and biochemical effects of itol A on the brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae).

Itol A, a novel isoryanodane diterpene derived from Itoa orientalis Hemsl., has potent activities against insect pests. This study was conducted to determine the contact toxicity and biochemical effects of itol A on the Nilaparvata lugens. After macropterous females of N. lugens were exposed to itol A from 0.5 to 24 h, the mortality and poisoning symptoms were measured. Effects of itol A on the major enzymes activity and oxidative stress level were assessed in dose-response (with LD10-LD70 at 24 h) and time-course (with LD50 at 0.5-24 h) experiments for the potential toxicity mechanisms. Based on the results, the mortality of N. lugens showed significant dose- and time-dependent effects, with the 24-h LD50 value was 0.58 µg/insect. The symptoms of excitation, convulsion and paralysis were also observed. However, acetylcholinesterases (AChE) activity was not altered after itol A treatment compared to control. Na+/K+-ATPases, Ca2+-ATPases, Ca2+/Mg2+-ATPases, glutathione S-transferases (GSTs), cytochrome P450 monooxygenases (P450s), superoxide dismutases (SOD) and catalases (CAT) activities were significantly reduced in dose-response and time-course experiments. While acid phosphatases (ACP) and glutathione peroxidases (GPX) activities were significantly increased. We further revealed that itol A exposure resulted in the decrease of GSH/GSSG (reduced to oxidized glutathione) ratio and the increase of hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels in both experiments. The results indicated that the inhibition of Na+/K+-ATPases, Ca2+-ATPases, Ca2+/Mg2+-ATPases, GSTs, P450s, SOD and CAT activities and the induction of oxidative stress was one of the potential biochemical mechanisms of itol A against N. lugens.

Using PhyloSuite for molecular phylogeny and tree-based analyses.

Phylogenetic analysis has entered the genomics (multilocus) era. For less experienced researchers, conquering the large number of software programs required for a multilocus-based phylogenetic reconstruction can be somewhat daunting and time-consuming. PhyloSuite, a software with a user-friendly GUI, was designed to make this process more accessible by integrating multiple software programs needed for multilocus and single-gene phylogenies and further streamlining the whole process. In this protocol, we aim to explain how to conduct each step of the phylogenetic pipeline and tree-based analyses in PhyloSuite. We also present a new version of PhyloSuite (v1.2.3), wherein we fixed some bugs, made some optimizations, and introduced some new functions, including a number of tree-based analyses, such as signal-to-noise calculation, saturation analysis, spurious species identification, and etc. The step-by-step protocol includes background information (i.e., what the step does), reasons (i.e., why do the step), and operations (i.e., how to do it). This protocol will help researchers quick-start their way through the multilocus phylogenetic analysis, especially those interested in conducting organelle-based analyses.

Itol A May Affect the Growth and Development of Spodoptera frugiperda through Hijacking JHBP and Impeding JH Transport.

Isoryanodane and ryanodane diterpenes have a carbon skeleton correlation in structures, and their natural product-oxidized diterpenes show antifeedant and insecticidal activities against Hemiptera and Lepidoptera. While ryanodine mainly acts on the ryanodine receptor (RyR), isoryanodane does not. In this study, we demonstrated that itol A, an isoryanodane diterpenoid, could significantly downregulate the expression level of juvenile hormone-binding protein (JHBP), which plays a vital role in JH transport. RNAi bioassay indicated that silencing the Spodoptera frugipreda JHBP (SfJHBP) gene decreased itol A activity, which confirmed the developmental phenotypic observation. Parallel reaction monitoring (PRM) further confirmed that itol A affected JHBP's expression abundance. Although JHBP is not proven as the direct or only target of itol A, we confirmed that itol A's action effect depends largely on JHBP and that JHBP is a potential target of itol A. We present foundational evidence that itol A inhibits the growth and development of Spodoptera frugiperda mainly through hijacking JHBP.

Effects of itol A on the larval growth and development of Spodoptera frugiperda (Lepidoptera: Noctuidae).

BACKGROUND: Itol A, extracted from Itoa orientalis Hemsl. (Flacourtiaceae), possesses bioactivity on Spodoptera litura (Lepidoptera: Noctuidae) and Nilaparvata lugens (Stål) (Hemiptera: Delphacidae). Our previous study showed that the effects on Spodoptera frugiperda, a destructive pest found worldwide, were similar to those of fenoxycarb (FC), a juvenile hormone analog. Thus, we speculate that itol A could have growth-regulating effects. The current work explored juvenile hormone (JH) levels and mRNA levels of crucial JH signaling pathway enzyme genes in S. frugiperda larvae treated with itol A and FC. RESULTS: Itol A caused severe growth obstacles in S. frugiperda, extended the larval duration and reduced the mean worm weight and body length rates. Three and 7 days after exposure to a sublethal concentration of itol A (500 mg L-1 ), the JH level of the larvae significantly decreased by 36.59% and 22.70%, respectively. qPCR inferred that the mRNA expression levels of crucial JH metabolism enzymes (SfJHE and SfJHEH) significantly increased by 6.58-fold and 2.12-fold, respectively, relative to the control group 3 days after treatment. CONCLUSIONS: Itol A adversely affects the development of S. frugiperda. We propose that this effect was caused by decreasing JH levels and disrupting the JH signaling pathway via mediating its synthetic and metabolic crucial enzymes. © 2021 Society of Chemical Industry.

Effects of botanical pesticide itol A against the tobacco cutworm, Spodoptera litura (Fab.).

Itol A, an isoryanodane diterpene derived from Itoa orientalis Hemsl. (Flacourtiaceae), is a potential plant-based insecticide. However, the effect of itol A on the tobacco cutworm [Spodoptera litura (Fab.) (Lepidoptera: Noctuidae)], an important and widely distributed insect pest, remains unclear. In this study, the toxicity and inhibitory potency of itol A on S. litura were evaluated. The results indicated that itol A exhibited larvicidal activity against the third instar larvae in a concentration-dependent manner (LC50 875.48 mg/L at 96 h). Antifeedant activity also was observed, and the 24-h AFC50 values were 562.05 and 81.47 mg/L in the no-choice and choice experiments, respectively. The insect growth was inhibited after treatment of itol A, as reflected by long developmental periods, low-quality pupae, and various abnormalities. Itol A exerted ovicidal effect on S. litura, with an estimated LC50 of 759.30 mg/L. Itol A deterred oviposition in the choice experiment (ODI50 909.60 mg/L). Besides, the activities of α-amylase, general protease, superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were inhibited after itol A treatment over time compared to controls, which may be a relevant mechanism underlying the toxicity of itol A toward S. litura. However, the activities of lipase, carboxylesterase (CarE), glutathione S-transferase (GST), and cytochrome P450 monooxygenase (P450) were increased. Taken together, these results suggest that itol A could be a good botanical pesticide to reduce the population of S. litura in integrated pest management programs.